Hugging Face's logo

Spaces:
debaghtk
/
patent-ai
Runtime error

App Files Community
patent-ai / data /4000042.html
debaghtk's picture
debaghtk
download 1000 patents
98c6817
raw
history blame contribute delete
69.9 kB
<!DOCTYPE HTML>
<html lang="en">
 <head>
 <meta http-equiv="Content-Type" content="text/html; charset=utf-8"/>
 <title>US Patent for Diagnostic reagent for the determination of amylase Patent (Patent # 4,000,042 issued December 28, 1976) - Justia Patents Search</title>
 <meta name="description" content="Disclosed herein is a diagnostic agent for determining the .alpha.-amylase content of a sample comprising a first compound selected from the group consisting of maltotetraose, maltopentaose, and maltohexaose; and a second component comprising an appropriate glucose detecting reagent, such as glucose oxidase or glucose dehydrogenase.">
 <meta name="keywords" content="Diagnostic reagent for the determination of amylase, Patent">
<meta name="robots" content="index, follow">
<link rel="canonical" href="https://patents.justia.com/patent/4000042" />
<link rel="stylesheet" type="text/css" href="https://justatic.com/v/20231003111152/shared/css/core.css">
<link href="https://plus.google.com/113923255404381000955" rel="publisher" />
<link rel="icon" type="image/svg+xml" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_144.svg" />
 <link rel="icon" type="image/png" sizes="144x144" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_144.png" />
 <link rel="icon" type="image/png" sizes="128x128" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_128.png" />
 <link rel="icon" type="image/x-icon" sizes="16x16 24x24 32x32 48x48 64x64 96x96 128x128" href="https://justatic.com/v/20231003111152/shared/images/justia/justia.ico" />
 <link rel="apple-touch-icon" type="image/png" sizes="57x57" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_57.png" />
 <link rel="apple-touch-icon" type="image/png" sizes="72x72" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_72.png" />
 <link rel="apple-touch-icon" type="image/png" sizes="114x114" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_114.png" />
 <link rel="apple-touch-icon" type="image/png" sizes="144x144" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_flat_144.png" />
 <link rel="apple-touch-icon-precomposed" type="image/png" sizes="57x57" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_3d_57.png" />
 <link rel="apple-touch-icon-precomposed" type="image/png" sizes="72x72" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_3d_72.png" />
 <link rel="apple-touch-icon-precomposed" type="image/png" sizes="114x114" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_3d_114.png" />
 <link rel="apple-touch-icon-precomposed" type="image/png" sizes="144x144" href="https://justatic.com/v/20231003111152/shared/images/justia/jbutton_3d_144.png" />
 <link rel="shortcut icon" type="image/vnd.microsoft.icon" sizes="16x16 24x24 32x32 48x48 64x64 96x96 128x128" href="https://justatic.com/v/20231003111152/shared/images/justia/justia.ico" />
 <meta name="viewport" content="width=device-width, initial-scale=1.0">
<meta name="trigger-ga" content="1" />
<!-- Google Tag Manager -->
 <script>(function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':
 new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],
 j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src=
 'https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);
 })(window,document,'script','dataLayer','GTM-P2QTGCR');</script>
 <!-- End Google Tag Manager -->
 </head>
<body class="patents sticky-footer content-right-sidebar" data-project="patents" data-directive="j-gdpr" data-privacy-policy-url="https://www.justia.com/privacy-policy/">
 <!-- Google Tag Manager (noscript) -->
 <noscript><iframe src="https://www.googletagmanager.com/ns.html?id=GTM-P2QTGCR"
 height="0" width="0" style="display:none;visibility:hidden"></iframe></noscript>
 <!-- End Google Tag Manager (noscript) -->
<header id="header" data-directive="j-topnav">
 <div id="sticky-nav">
 <div class="burger-wrapper">
 <div id="hamburger" data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header" data-gtm-label="Hamburger" class="hamburger">
 <div class="inner-ham" style="pointer-events:none">
 <span class="lns ln2-1" style="pointer-events:none"></span>
 <span class="lns ln2-2" style="pointer-events:none"></span>
 </div>
 </div>
 <div id="header-nav">
 <div class="inner-wrapper">
 <div class="logo-wrapper">
 <a href="/" title="Justia" class="logo">
 <img data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header" data-gtm-label="Logo" src="https://justatic.com/v/20231003111152//shared/images/logos/justia.svg" alt="Justia" onerror="this.onerror=null; this.src='https://justatic.com/v/20231003111152//shared/images/logos/justia.png">
 <img data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header" data-gtm-label="Logo - Subdomain" class="sub-domain" src="https://justatic.com/v/20231003111152//shared/images/logos/justia-patents.svg" alt="Dockets" onerror="this.onerror=null; this.src='https://justatic.com/v/20231003111152//shared/images/logos/justia-patents.png'">
 </a>
 </div>
 <div class="buttons-wrapper">
 <div class="topnav-button search-icon" id="search-icon-button">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/white/search.svg" alt="search" width="22" height="22"/>
 </div>
 <div class="topnav-button sign-in-icon" id="sign-in-icon-button">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/white/profile.svg" alt="My Account" width="20" height="20">
 </div>
 <div id="search-form-wrapper">
 <form action="/search" method="get" class="search-form jform" id="search-form">
 <div class="input-wrapper">
 <input type="text" name="q" value="" id="querybox" title="Search" placeholder="Search">
 </div>
 <button type="submit">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/search.svg" alt="search" width="22" height="22"/>
 </button>
 </form>
 </div>
 <div id="accounts-buttons-wrapper" data-directive="j-sign-in-accounts">
 <span class="topnav-button accounts-buttons login-buttons" id="login-buttons-wrapper">
 <span class="login-buttons-wrapper">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Account" data-gtm-label="Log In" data-directive="j-login" href="https://accounts.justia.com/signin?destination=https%3A%2F%2Fpatents.justia.com%2Fpatent%2F4000042" class="login">Log In</a>
 <a href="https://accounts.justia.com/" class="sign-up">Sign Up</a>
 </span>
 </span>
 </div>
 </div>
 </div>
 </div>
 </div>
 </div>
 <div id="subnavigation">
 <div class="subnavigation-wrapper">
 <ul>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Find a Lawyer" href="https://www.justia.com/lawyers">Find a Lawyer</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Ask a Lawyer" href="https://answers.justia.com/">Ask a Lawyer</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Research the Law" href="https://www.justia.com/">Research the Law</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Law Schools" href="https://www.justia.com/law-schools/">Law Schools</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Laws &amp; Regs" href="https://law.justia.com/">Laws &amp; Regs</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Newsletters" href="https://connect.justia.com/case-law-summary-newsletters">Newsletters</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Marketing Solutions" href="https://www.justia.com/marketing/">Marketing Solutions</a>
 <div class="sublist">
 <div class="sublist-section">
 <ul>
 <li class="sublist-section-title"><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Justia Connect" href="https://www.justia.com/marketing/connect/">Justia Connect</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Pro Membership" href="https://www.justia.com/marketing/connect/#pro-membership">Pro Membership</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Basic Membership" href="https://www.justia.com/marketing/connect/#basic-membership">Basic Membership</a></li>
 </ul>
 </div>
 <div class="sublist-section">
 <ul>
 <li class="sublist-section-title"><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Justia Lawyer Directory" href="https://www.justia.com/marketing/lawyer-directory/">Justia Lawyer Directory</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Platinum Placements" href="https://www.justia.com/marketing/lawyer-directory/platinum/">Platinum Placements</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Gold Placements" href="https://www.justia.com/marketing/lawyer-directory/gold/">Gold Placements</a></li>
 </ul>
 </div>
 <div class="sublist-section">
 <ul>
 <li class="sublist-section-title"><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Justia Elevate" href="https://www.justia.com/marketing/elevate/">Justia Elevate</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="SEO" href="https://www.justia.com/marketing/elevate/search-engine-optimization/">SEO</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Websites &amp; LSA Management" href="https://www.justia.com/marketing/elevate/law-firm-websites/">Websites</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Blogs" href="https://www.justia.com/marketing/elevate/law-blogs/">Blogs</a></li>
 </ul>
 </div>
 <div class="sublist-section">
 <ul>
 <li class="sublist-section-title"><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Justia Amplify" href="https://www.justia.com/marketing/amplify/">Justia Amplify</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="PPC Management" href="https://www.justia.com/marketing/amplify/cpc-ppc/">PPC Management</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Google Business Profile" href="https://www.justia.com/marketing/amplify/google-business-profile/">Google Business Profile</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Social Media" href="https://www.justia.com/marketing/amplify/social-media-networking/">Social Media</a></li>
 </ul>
 </div>
 <div class="sublist-section">
 <ul>
 <li class="sublist-section-title"><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Header Navigation" data-gtm-label="Justia Onward Blog" href="https://onward.justia.com/">Justia Onward Blog</a></li>
 </ul>
 </div>
 </div>
 </li>
 </ul>
 </div>
 </div>
 </header>
 <div class="has-topnav-padding font-opensans" data-directive="j-announcement" data-project="patents"></div>
 <div id="main-content" class="main-content font-opensans">
 <nav class="breadcrumbs font-helvetica small-font">
 <a href="https://www.justia.com">Justia</a>
 <span class="breadcrumb-separator"></span>
 <a href="/">Patents</a> <span class="breadcrumb-separator"></span>
 US Patent for Diagnostic reagent for the determination of amylase Patent (Patent # 4,000,042)
 </nav>
 <div class="primary-content-wrapper">
 <div class="primary-content">
 <ins class="adsbygoogle"
 style="display:block"
 data-ad-client="ca-pub-4319265774032289"
 data-ad-slot="2491452856"
 data-ad-format="auto"></ins>
 <div class="wrapper jcard has-padding-30 blocks">
 <div class="wrapper">
 <div class="title-wrapper">
 <h1 class="heading-1"> Diagnostic reagent for the determination of amylase </h1>
 </div>
 <div class="wrap">
 <div id="byline" class="small-font">
 <time datetime="1975-01-20" class="color-dove-gray">Jan 20, 1975</time>
- <a href="/assignee/e-i-du-pont-de-nemours-and-company">E. I. Du Pont de Nemours and Company</a>
</div> <div id="abstract" class="to-xlarge-font">
 <p>Disclosed herein is a diagnostic agent for determining the .alpha.-amylase content of a sample comprising a first compound selected from the group consisting of maltotetraose, maltopentaose, and maltohexaose; and a second component comprising an appropriate glucose detecting reagent, such as glucose oxidase or glucose dehydrogenase.</p>
 </div>
 <div id="company-latest" class="wrapper">
 <div class="title-wrapper">
 <h2 class="heading-2">Latest E. I. Du Pont de Nemours and Company Patents:</h2>
 </div>
 <div class="wrap indented">
 <ul class="list-gutter--tiny">
 <li>
 <a href="/patent/20230279413">PLANT GENOME MODIFICATION USING GUIDE RNA/CAS ENDONUCLEASE SYSTEMS AND METHODS OF USE</a>
 </li>
 <li>
 <a href="/patent/20230235342">METHODS FOR PRODUCING A COMPLEX TRANSGENIC TRAIT LOCUS</a>
 </li>
 <li>
 <a href="/patent/20230235345">PLANT GENOME MODIFICATION USING GUIDE RNA/CAS ENDONUCLEASE SYSTEMS AND METHODS OF USE</a>
 </li>
 <li>
 <a href="/patent/20230220407">MAIZE EVENT DP-004114-3 AND METHODS FOR DETECTION THEREOF</a>
 </li>
 <li>
 <a href="/patent/20230212595">GENERATION OF SITE SPECIFIC INTEGRATION SITES FOR COMPLEX TRAIT LOCI IN CORN AND SOYBEAN, AND METHODS OF USE</a>
 </li>
 </ul>
 </div>
 </div>
 <div id="content-navigation">
 <span class="right-10">Skip to:</span>
 <span>
 <a href="#description">Description</a>
 </span>
 <span>&nbsp;·&nbsp;</span><span>
 <a href="#claims">Claims</a>
 </span>
 <span>&nbsp;·&nbsp;</span><span>
 <a href="#citations">References Cited</a>
 </span>
 <span>&nbsp;·&nbsp;</span><span><a href="#history">Patent History</a></span>
 <span>&nbsp;·&nbsp;</span><span>
 <a href="#history">Patent History</a>
 </span>
 </div>
 <div id="description" class="wrapper">
 <div class="title-wrapper">
 <strong class="heading-2">Description</strong>
 </div>
 <div class="wrap">
 <strong class="heading-4">BACKGROUND OF THE INVENTION</strong>
<p>1. Field of the Invention</p>
<p>This invention relates to a procedure for the qualitative determination of .alpha.-amylase in a sample, and more particularly to a diagnostic agent for the enzymatic determination of .alpha.-amylase. The qualitative measurement of .alpha.-amylase concentrations in a sample is useful in medical diagnostics.</p>
<p>2. Discussion of the Prior Art</p>
<p>The enzyme .alpha.-amylase is to be found in salivary and pancreatic juices. It is well known that this enzyme reacts with various carbohydrates, particularly starch, to form maltose, and that the enzyme maltase will react with maltose to form glucose. These two reactions form the basis for intestinal digestion. See, for example, Biochemistry by Cantarow and Schepartz, page 263.</p>
<p>It is also well known that the presence of glucose can be determined by various coupled reactions. This knowledge has led to several proposed methods for measuring amylase activity in conjunction with maltose by utilizing its ability to convert starch to glucose. See, for example, the article by H. W. Schiwara in Arztl Chab. 17:340-343 (1971) and the paper entitled Columetric Rate Determination of Serum Amylase Activity given at the National Symposium on Enzyme Chemistry in 1972.</p>
<p>Techniques involving starch, however, do not allow truly quantitative amylase determinations. The amylase reacts with each of the constituents of starch to produce different reaction products, and the kinetics are unpredictable. This is particularly troublesome in rate determinations where the measurements are made within a short span of time, during which the reaction kinetics are even less predictable. As a result of this, measurements made using starch as the substrate for .alpha.-amylase activity do not, and in fact cannot, give a true measure of .alpha.-amylase concentrations in the test sample.</p>
<p>Furthermore, the test proceedings of the prior art are not capable of differentiating between salivary .alpha.-amylase and pancreatic .alpha.-amylase.</p>
<p>It is an object of the present invention to provide a diagnostic agent for the determination of .alpha.-amylase which allows quantitative measurements to be made. It is a further object of this invention to provide a diagnostic agent which can be used in a rate determination process for the determination of amylase so that a rapid quantitative measurement of the amount of amylase present in a sample can be made.</p>
<strong class="heading-4">SUMMARY OF THE INVENTION</strong>
<p>These and other objects are accomplished by a diagnostic agent for the determination of .alpha.-amylase comprising a first component selected from the group consisting of maltotetraose, maltopentaose, or maltohexaose; and a second component comprising an appropriate glucose detecting reagent. Preferably, the agent also comprises maltase.</p>
<p>The use of maltotetraose or maltopentaose as the substrate produces a truly stoichiometric determination when maltohexaose is used as the substrate, however, is believed to be slight and, therefore, tolerable.</p>
<p>The reaction between pancreatic .alpha.-amylase and substrate will produce maltaose, maltotriose and glucose. The glucose evolving from the reaction between the pancreatic .alpha.-amylase and the substrate can be measured by any one of a number of well known techniques which will be discussed below. Salivary .alpha.-amylase, however, does not produce glucose. The reaction between the salivary .alpha.-amylase and the substrate appears to produce only maltose. If the reaction solution contained a maltase, preferably .alpha.-glucosidase, the maltose and maltotriose produced by the reaction between the substrate and the .alpha.-amylase will be converted to glucose and the glucose can be measured by any one of a number of conventional techniques. The reaction proceeds most favorably at a temperature of between about 20.degree. and 50.degree. C. and at a pH of between about 5 and 10.</p>
<p>Since the only two sources of .alpha.-amylase are the pancreas and the salivary glands, total .alpha.-amylase can be determined by a solution containing both the substrate and the maltase; pancreatic .alpha.-amylase can be determined by a solution from which the maltase is absent, and salivary .alpha.-amylase can be determined by taking the difference between the total and the pancreatic .alpha.-amylase.</p>
<p>In the preferred embodiment, the substrate is maltopentaose and the step of determining the glucose evolved is accomplished by measuring the rate at which glucose is evolved. In a still more-preferred embodiment .beta.-glycerol phosphate is used as a buffer. Any other conventional buffer which will not interfere with the ability of the .alpha.-glucosidase to convert maltose into glucose can also be used.</p>
<p>There are a number of ways in which glucose can be determined. Any one of these ways can be used. A particularly convenient approach is to use a coupled enzyme determination for glucose. One suitable pair of coupled enzyme reactions are defined by the following: ##EQU1## where ATP is adenosine triphosphate, ADP is adenosine diphosphate, NAD is .beta.-nicotinamide-adenine dinucleotide, NADH is the reduced form of .beta.-nicotinamide-adenine dinucleotide, NADH is G-6-PDH is glucose-6-phosphate dehydrogenase. Since the reduced form of .beta.-nicotinamide-adenine dinucleotide absorbs light very strongly at 340 millimicrons, while the oxidized form does not, the rate at which NADH is evolved is directly proportional to the increase in absorbance of light at 340 millimicrons at constant temperature, usually 15.degree. C. to 50.degree. C, and constant pH. This increase can readily be measured by those skilled in the art, using a conventional spectrophotometer. Since the rate of formation of NADH is proportional to the rate at which glucose is evolved, the increase in absorbance at 340 millimicrons can be used as a direct measure of the original concentration of .alpha.-amylase in the sample. It should also be understood that other wavelengths, e.g., 366 millimicrons, can also be used for the foregoing purpose.</p>
<p>Another pair of coupled enzyme reactions which can be used to determine the presence of glucose are the following: ##EQU2## Other coupled enzyme reactions can also be used to measure the glucose content, or the glucose content may be measured directly.</p>
<p>Although .alpha.-glucosidase has been mentioned as the preferred maltase for use in the present invention, it should be noted that other forms of maltase can be used in the present invention. In the preferred embodiment, the substrate plus the maltase, if maltase is used, are added to the solution last. This is so that any glucose present in the sample will be used up by the other materials, prior to the introduction of the substrate. By this method, any glucose that is measured, after the substrate has been added to the solution, is due to the presence of .alpha.-amylase in the sample. The invention will be described with reference to the following examples, and FIG. 1 which is a plot of the change in absorbance .DELTA.A per minute, in multiabsorbance units, of the reaction solution as a function of the amylase concentration in the solution for Example 1 and 2.</p>
<strong class="heading-4">BRIEF DESCRIPTION OF THE FIGURE</strong>
<p>The FIGURE shows the change in absorbance over a 5 minute period (in .DELTA.A/minute units) for a standard solution of various amylase activity (in Somogyi Units/dl) as described in Examples 1 and 2.</p>
<strong class="heading-4">EXAMPLE 1</strong>
<p>An amylase activity standard curve was generated on a Cary 14 spectrophotometer by following the rate of change in the absorbance of NADH. The sample used for the standard curve solution was a 0.85 mg/ml solution of partially purified human pancreatic amylase dissolved in human serum. This resulted in a sample with an amylase activity of 1600 Somogyi units/dl. Dilutions of this serum were made with a 0.85% saline solution to achieve lower value.</p>
<p>The following solutions were used in the assay.</p>
<p>.beta.-gylcerol phosphate buffer, 0.06 M, pH 6.8</p>
<p>Nad, 75 mg/ml H.sub.2 O</p>
<p>Atp, 30 mg/ml H.sub.2 O</p>
<p>NaCl Mg SO.sub.4, 50 mg MgSO.sub.4 + 30 mg NaCl/ml H.sub.2 O</p>
<p>Hexokinase, 2456 IU/ml</p>
<p>Glucose-6-phosphate dehydrogenase, 1000 IU/ml</p>
<p>maltotetraose, 200 mg/ml H.sub.2 O</p>
<p>.alpha.-glucosidase, 10 mg/ml</p>
<p>The assay was performed as follows: To 4.50 milliliters of .beta.-glycerol phosphate buffer, 0.1 milliliters of serum were added and allowed to equilibriate at 37.degree. C. for 1.5 minutes. The following reagents were then added:</p>
<p>Nad, 0.1 ml (7.5 mg), ATP, 0.1 ml (3.0 mg)</p>
<p>NaCl--MgSO.sub.4 - 0.05 ml (1.5 mg & 2.5 mg)</p>
<p>Hexokinase, 0.01 ml (25 IU)</p>
<p>Glucose-6-phosphate dehydrogenase, 0.01 ml (10 IU)</p>
<p>the reagents were mixed and the glucose in the sample was consumed in a 2 minute period. The reaction was then initiated by adding 0.04 ml .alpha.-glucosidase (10 IU) and 0.1 ml maltotetraose (20 mg). An increase in absorbance at 340 nm was monitored for 5 minutes. The change in absorbance over the 5 minute period for various solutions of the standard sample is shown by the open circles and the solid line in the figure.</p>
<strong class="heading-4">EXAMPLE 2</strong>
<p>The amylase detection system has also been adapted for automatic use by the Du Pont Automatic Clinical Analyzer (aca). Reagents for the automatic determination were prepared as described below:</p>
<p>1. A blend containing maltotetraose, ATP, polyethylene glycol (PEG) 2000 and .beta.-glycerol phosphate was prepared in the following proportions: 8 mg maltotetraose; 4.8 mg ATP; 7.2 mg PEG; 5.5 mg .beta.-glycerol phosphate and 69.4 mg mannitol. This blend was tableted producing tablets which contained 4.4 mg maltotetraose, 2.2 mg ATP.</p>
<p>2. A tablet was prepared containing 8.3 mg NAD, 1.9 mg MgSO.sub.4, 4.1 mg PEG and 85 mg .beta.-glycerol phosphate.</p>
<p>3. .alpha.-glucosidase, hexokinase and glucose-6-phosphate dehydrogenase were combined in a 50% glycerol albumin solution with 20 IU hexokinase, 18 IU glucose-6-phosphate dehydrogenase and 20 IU .alpha.-glucosidase used per test.</p>
<p>Analytical test packs for the aca were assembled containing two NAD, MgSO.sub.4, .beta.-glycerol phosphate buffer tablets, one maltotetraose, ATP tablet and 0.050 milliliters of the enzyme solution. These reagents were placed in the reagent compartments of a test pack such as that described in U.S. Pat. No. 3,476,515.</p>
<p>The aca was programmed to use 0.100 ml. of sample, 4.9 ml of water, and to report the results of the assay in milliabsorbance units/minute. The sample used was similar to that used in Example 1 except that the sample had an amylase activity of 750 Somogyi units/dl. Detection of this sample were also made as disclosed in Example 1.</p>
<p>The results of tests on various dilutions of the sample are represented by the solid clots and the dashed line in the figure.</p>
<strong class="heading-4">EXAMPLE 3</strong>
<p>Comparisons of the relative specificity of maltotetraose were made. Purified human pancreatic .alpha.-amylase and human salivary .alpha.-amylase were compared as to their abilities to react with the substrate maltotetraose. In the presence of the complete coupling system, .alpha.-glucosidase, ATP, NAD and G-6-PDH, both amylases appeared to react similarly. However, in the absence of the .alpha.-glucosidase, the salivary enzyme did not produce any glucose. This means that the salivary enzyme splits the maltotetraose into two maltose molecules only. The pancreatic enzyme produces a significant amount of glucose by its action on the maltotetraose directly. Thus, the method provides a means to determine the salivary and pancreatic amylase contributions to the gross rate, by running an additional assay in the absence of .alpha.-glucosidase.</p>
<strong class="heading-4">EXAMPLE 4</strong>
<p>The specificity of maltopentaose was compared with that of maltotetraose using the following reagents:</p>
<p>2 NAD, MgSO.sub.4, .beta.-glycerol phosphate buffer tablets (Of Example 2)</p>
<p>3 mg ATP</p>
<p>5 mg maltotetraose or 2.2 mg maltopentose</p>
<p>25 units hexokinase</p>
<p>28 units glucose-6-phosphate dehydrogenase</p>
<p>7 units .alpha.-glucosidase</p>
<p>Tablets were dissolved in 4.90 ml water and the reagents listed above were added to this solution. 0.020 milliliters of the sample of Example 1 were added to this solution. A blank reagent was run in the absence of sample and the change in absorbance (.DELTA.A) was measured at 340 nm after a 4 minute incubation in a Cary 14 spectrometer at 37.degree. C. The results were as follows:</p>
<p>______________________________________ Substrate .DELTA.A/min. ______________________________________ test maltotetraose .078 blank .026 test maltopentaose .101 blank .0015 ______________________________________</p>
<p>The results show that the observed rate with the same sample is higher for maltopentaose than for maltotetraose. In addition, the blank rate, i.e., the reaction of the .alpha.-glucosidase with substrate is much higher with maltotetraose and is less desirable. Thus, maltopentaose would appear to be highly sensitive and would eliminate the need for closely monitoring the extent of the blank reaction.</p>
</div>
 </div>
 <div id="claims" class="wrapper">
 <div class="title-wrapper">
 <h2 class="heading-2">Claims</h2>
 </div>
 <div class="wrap">
 <div class="claim">
 <p> 1. Diagnostic agent for the determination of.alpha.-amylase in a body fluid comprising: </p><dl><dd> a. a first component selected from the group of reagents consisting of maltotetraose, maltopentaose and maltohexaose; and </dd><dd> b. a second component comprising an enzymatic glucose detecting reagent. </dd></dl>
 </div>
 <div class="claim">
 <p> 2. The agent of claim 1 wherein said second component comprises a glucose detecting reagent selected from the group consisting of glucose oxidase and glucose dehydrogenase. </p>
 </div>
 <div class="claim">
 <p> 3. Diagnostic agent for the determination of.alpha.-amylase in a body fluid comprising: </p><dl><dd> a. maltotetraose; and </dd><dd> b. a second component comprising an enzymatic glucose detecting reagent. </dd></dl>
 </div>
 <div class="claim">
 <p> 4. Diagnostic agent for the determination of.alpha.-amylase in a body fluid comprising: </p><dl><dd> a. maltopentaose; and </dd><dd> b. a second component comprising an enzymatic glucose detecting reagent. </dd></dl>
 </div>
 <div class="claim">
 <p> 5. Diagnostic reagents for the determination of salivary.alpha.-amylase in a body fluid comprising: </p><dl><dd> a. a first component selected from the group of reagents consisting of maltotetraose, maltopentaose, and maltohexaose; </dd><dd> b. a second component comprising maltase; and </dd><dd> c. a third component comprising an enzymatic glucose detecting reagent. </dd></dl>
 </div>
 <div class="claim">
 <p> 6. The agent of claim 5 wherein said first component is maltotetraose. </p>
 </div>
 <div class="claim">
 <p> 7. The agent of claim 5 wherein said first component is maltopentaose. </p>
 </div>
 <div class="claim">
 <p> 8. The agent of claim 5 wherein said maltase is.alpha.-glucosidase. </p>
 </div>
 <div class="claim">
 <p> 9. The agent of claim 5 wherein said second component comprises a glucose detecting reagent selected from the group consisting of glucose oxidase and glucose dehydrogenase. </p>
 </div>
 <div class="claim">
 <p> 10. The agent of claim 1 further comprising.alpha.-glucosidase. </p>
 </div>
 <div class="claim">
 <p> 11. The agent of claim 5, further comprising.alpha.-glucosidase. </p>
 </div>
 </div>
 </div>
 <div id="citations" class="wrapper">
 <div class="title-wrapper">
 <strong class="heading-2">Referenced Cited</strong>
 </div>
 <div class="wrap">
 <strong class="heading-4">U.S. Patent Documents</strong>
<div class="wrap block">
 <table id="us-citations" class="zebra -zb table-full-width">
 <tbody class="table-bordered">
 <tr class="zebra -zb table-padding-full">
 <td class="number">
 3778350
 </td>
 <td class="date">
 December 1973
 </td>
 <td class="inventor">
 Bergmeyer
 </td>
 </tr>
 </tbody>
 </table>
</div>
 <strong class="heading-4">Other references</strong>
 <ul>
 <li>Greenwaad et al., Die Starke, "Studies on Starch Degrading Enzymes -- Part VIII", 20(5), 1968, pp. 139-150.
Okada et al., Agr. Biol. Chem., vol. 33, No. 6, (1969), pp. 900-906.</li>
 </ul>
 </div>
 </div>
 <div id="history" class="wrapper">
 <div class="title-wrapper">
 <strong class="heading-2">Patent History</strong>
 </div>
 <div class="wrap">
 <strong>Patent number</strong>: 4000042<br>
<strong>Type: </strong>
 Grant
<br>
<strong>Filed</strong>: Jan 20, 1975<br>
<strong>Date of Patent</strong>: Dec 28, 1976<br>
<strong>Assignee</strong>:
<a href="/assignee/e-i-du-pont-de-nemours-and-company">E. I. Du Pont de Nemours and Company</a>
(Wilmington, DE)
 <br>
<strong>Inventor</strong>:
<a href="/inventor/thomas-h-adams">Thomas H. Adams</a>
(Mission Viejo, CA)
 <br>
<strong>Primary Examiner</strong>: <a href="/examiner/a-louis-monacell">A. Louis Monacell</a>
 <br>
<strong>Assistant Examiner</strong>: <a href="/examiner/esther-l-massung">Esther L. Massung</a>
 <br>
<strong>Application Number</strong>: 5/542,236<br>
 </div>
</div>
 <div id="classifications" class="wrapper">
 <div class="title-wrapper">
 <strong class="heading-2">Classifications</strong>
 </div>
 <div class="wrap">
<strong>Current U.S. Class</strong>:
<strong>
 195/1035R;
 </strong>
 195/99;
 195/1035C;
 195/63
 <br>
<strong>International Classification</strong>:
C12K 100;
 G01N 3114;
 <br>
</div>
 </div>
 </div>
 </div>
 </div>
 </div>
 <div class="sidebar primary-sidebar">
 <aside
 data-directive="j-lead"
 data-type="cornell"
 data-impression="0">
 </aside>
 <aside class="jlead-v2-wrapper jcard -loading" data-directive="j-dynamic-lead" data-lead-id="146">
 <div class="jspinner -double-bounce"><div class="-crl -crl1 bg-justia-blue"></div><div class="-crl -crl2 bg-justia-blue"></div></div>
</aside>
 <aside id="ask-a-lawyer-widget" class="jcard has-padding-content-block-30 block">
 <p class="heading-4">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Sidebar" data-gtm-label="All - Ask a Lawyer" href="https://answers.justia.com/">Ask a Lawyer</a>
 </p>
 <div class="content">
 <div class="block-wrapper small-font" data-directive="j-ask-a-lawyer">
 <form data-gtm-type="submission" data-gtm-label="Ask a Lawyer" data-gtm-category="Widgets" name="ask-a-lawyer-form" method="post" action="https://answers.justia.com/ask" id="homepage-ask" accept-charset="utf-8">
 <div class="jfld-group">
 <strong>Question:</strong>
 <textarea name="question" class="jfld-block" id="ask-a-lawyer-textarea" rows="5" title="Please ask your question here and get free answers from lawyers." placeholder="Please ask your question here and get free answers from lawyers."></textarea>
 </div>
 <span class="block">
 <a id="answers-add-details">Add details</a>
 <div class="pull-right" id="char-court">120</div>
 </span>
 <div class="question-details clear-both hidden-content" id="question-details">
 <div class="jfld-group">
 <strong>Additional Details: </strong>
 <textarea name="details" id="ask-a-lawyer-details-textarea" rows="5" class="jfld-block" title="If you have additional details about your question fill them here" placeholder="If you have additional details about your question fill them here"></textarea>
 </div>
 <span class="block">
 <div class="pull-right" id="char-court-details">
 1000
 </div>
 </span>
 </div>
 <button type="submit" name="submit" class="pull-right button button-primary uppercase">
 Ask Question
 </button>
 </form>
 </div>
 </div>
</aside>
 <aside class="jcard jlead-v2-wrapper -loading" data-directive="j-dynamic-lead" data-property="patents">
 <div class="jspinner -double-bounce">
 <div class="-crl -crl1 bg-justia-blue"></div>
 <div class="-crl -crl2 bg-justia-blue"></div>
 </div>
</aside>
 <aside class="jcard has-padding-content-block-30 block">
 <p class="heading-4">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Sidebar" data-gtm-label="All - Find a Lawyer" href="//www.justia.com/lawyers/patents/locate" id="findalawyerlink">Find a Lawyer</a>
 </p>
 <div class="block-wrapper">
 <form data-gtm-type="submission" data-gtm-label="Find a Lawyer" data-gtm-category="Widgets" class="jform small-font jform--find-lawyer" name="find-a-lawyer-form" action="//lawyers.justia.com/search" method="get" accept-charset="utf-8">
 <div class="jfld-group">
 <input type="text" name="query" value="Patents" class="jfld-block" id="lawyer_practice_field" title="Legal Issue or Lawyer Name" placeholder="Legal Issue or Lawyer Name"/>
 </div>
 <div class="jfld-group -side-button clearfix">
 <input type="text" name="location" id="lawyer-location" title="City, State" placeholder="City, State" />
 <input type="submit" value="Search" class="button button-primary search uppercase" />
 </div>
 </form>
 <div class="text-center clear-both">
 <strong class="heading-6 has-no-bottom-margin">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Sidebar" data-gtm-label="All - Lawyers - Get Listed Now!" href="https://lawyers.justia.com/lawyer-directory-listings" class="button button-red-ghost">Lawyers - Get Listed Now!</a>
 </strong>
 <span class="tiny-font">Get a free directory profile listing</span>
 </div>
 </div>
</aside>
 <aside data-directive="j-find-a-lawyer" data-practice="patents" data-level="300"></aside>
 </div>
 </div>
 </div>
 <div id="footer" class="-v2 short bg-white color-black">
 <div class="wrapper has-no-padding">
 <div class="resources font-opensans has-padding-content-block-x2large clearfix">
 <div class="section-title">Justia Legal Resources</div>
 <div class="columns">
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Find a Lawyer" href="https://www.justia.com/lawyers">Find a Lawyer</a>
 <span class="expand" data-directive="j-expand" data-section="footer-01">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-01" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Bankruptcy Lawyers" href="https://www.justia.com/lawyers/bankruptcy">Bankruptcy Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Business Lawyers" href="https://www.justia.com/lawyers/business-law">Business Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Criminal Lawyers" href="https://www.justia.com/lawyers/criminal-law">Criminal Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Employment Lawyers" href="https://www.justia.com/lawyers/employment-law">Employment Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Estate Planning Lawyers" href="https://www.justia.com/lawyers/estate-planning">Estate Planning Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Family Lawyers" href="https://www.justia.com/lawyers/family-law">Family Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Personal Injury Lawyers" href="https://www.justia.com/lawyers/personal-injury">Personal Injury Lawyers</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Find a Lawyer - More" href="https://www.justia.com/lawyers">More... </a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Individuals" href="https://www.justia.com/individuals/">Individuals</a>
 <span class="expand" data-directive="j-expand" data-section="footer-02">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-02" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Bankruptcy" href="https://www.justia.com/bankruptcy/">Bankruptcy</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Criminal" href="https://www.justia.com/criminal/">Criminal</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Divorce" href="https://www.justia.com/family/divorce/">Divorce</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="DUI" href="https://www.justia.com/criminal/drunk-driving-dui-dwi/">DUI</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Estate Planning" href="https://www.justia.com/estate-planning/">Estate Planning</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Family Law" href="https://www.justia.com/family/">Family Law</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Personal Injury" href="https://www.justia.com/injury/">Personal Injury</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Individuals - More" href="https://www.justia.com/individuals/">More...</a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Business" href="https://www.justia.com/business/">Business</a>
 <span class="expand" data-directive="j-expand" data-section="footer-03">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-03" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Business Formation" href="https://www.justia.com/business-operations/starting-your-own-business/">Business Formation</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Business Operations" href="https://www.justia.com/business-operations/">Business Operations</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Employment" href="https://www.justia.com/employment/">Employment</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Intellectual Property" href="https://www.justia.com/intellectual-property/">Intellectual Property</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="International Trade" href="https://www.justia.com/international-law/international-trade-law/">International Trade</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Real Estate" href="https://www.justia.com/real-estate/">Real Estate</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Tax Law" href="https://www.justia.com/tax/">Tax Law</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Business - More" href="https://www.justia.com/business/">More... </a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Law Students" href="https://www.justia.com/law-students/">Law Students</a>
 <span class="expand" data-directive="j-expand" data-section="footer-04">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-04" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Law Schools" href="https://www.justia.com/law-schools/">Law Schools</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Admissions" href="https://www.justia.com/law-students/">Admissions</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Financial Aid" href="https://www.justia.com/law-students/">Financial Aid</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Course Outlines" href="https://www.justia.com/law-schools/outlines/">Course Outlines</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Law Journals" href="https://www.justia.com/law-schools/">Law Journals</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Blogs" href="https://blawgsearch.justia.com/blogs/categories/law-student">Blogs</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Employment" href="https://www.justia.com/law-students/">Employment</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Law Students - More" href="https://www.justia.com/law-students/">More...</a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="US Federal Law" href="https://law.justia.com/us/">US Federal Law</a>
 <span class="expand" data-directive="j-expand" data-section="footer-05">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-05" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="US Constitution" href="https://law.justia.com/constitution/us/">US Constitution</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="US Code" href="https://law.justia.com/codes/us">US Code</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Regulations" href="https://regulations.justia.com/">Regulations</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Supreme Court" href="https://supreme.justia.com/">Supreme Court</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Circuit Courts" href="https://law.justia.com/cases/federal/appellate-courts/">Circuit Courts</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="District Courts" href="https://law.justia.com/cases/federal/district-courts/">District Courts</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Dockets &amp; Filings" href="https://dockets.justia.com/">Dockets &amp; Filings</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="US Federal Law - More" href="https://law.justia.com/us/">More...</a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="US State Law" href="https://law.justia.com/us-states/">US State Law</a>
 <span class="expand" data-directive="j-expand" data-section="footer-06">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-06" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="State Constitutions" href="https://law.justia.com/us-states/">State Constitutions</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="State Codes" href="https://law.justia.com/codes/">State Codes</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="State Case Law" href="https://law.justia.com/cases/">State Case Law</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="California" href="https://law.justia.com/california/">California</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Florida" href="https://law.justia.com/florida/">Florida</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="New York" href="https://law.justia.com/new-york/">New York</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Texas" href="https://law.justia.com/texas/">Texas</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="US State Law - More" href="https://law.justia.com/us-states/">More... </a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Other Databases" href="https://www.justia.com/">Other Databases</a>
 <span class="expand" data-directive="j-expand" data-section="footer-07">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-07" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="COVID-19 Resources" href="https://www.justia.com/covid-19/">COVID-19 Resources</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Legal Blogs" href="https://blawgsearch.justia.com/">Legal Blogs</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Business Forms" href="https://contracts.justia.com/">Business Forms</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Product Recalls" href="https://recalls.justia.com/">Product Recalls</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Patents" href="https://patents.justia.com/">Patents</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Trademarks" href="https://trademarks.justia.com/">Trademarks</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Countries" href="https://www.justia.com/countries/">Countries</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Other Databases - More" href="https://www.justia.com/">More...</a></li>
 </ul>
 </div>
 <div class="footer-block">
 <div class="title">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Marketing Solutions" href="https://www.justia.com/marketing/">Marketing Solutions</a>
 <span class="expand" data-directive="j-expand" data-section="footer-08">
 <img src="https://justatic.com/v/20231003111152/shared/images/icons/svgfiles/blue/down.svg" alt="Arrow">
 </span>
 </div>
 <ul id="footer-08" class="footer-list -collapsed">
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Justia Connect Membership" href="https://www.justia.com/marketing/connect/">Justia Connect Membership</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Justia Lawyer Directory" href="https://www.justia.com/marketing/lawyer-directory/">Justia Lawyer Directory</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Justia Premium Placements" href="https://www.justia.com/marketing/lawyer-directory/#premium-placements">Justia Premium Placements</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Justia Elevate" href="https://www.justia.com/marketing/elevate/">Justia Elevate (SEO, Websites)</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Justia Amplify" href="https://www.justia.com/marketing/amplify/">Justia Amplify (PPC, GBP)</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Justia Onward Blog" href="https://onward.justia.com/">Justia Onward Blog</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Testimonials" href="https://www.justia.com/marketing/testimonials/">Testimonials</a></li>
 <li><a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer Resources" data-gtm-label="Marketing Solutions - More" href="https://www.justia.com/marketing/">More...</a></li>
 </ul>
 </div>
 </div>
 </div>
 </div>
 <div class="bg-justia-blue">
 <div class="wrapper">
 <div class="copyright">
 <span class="site">
 <span class="social-media">
 <a href="https://www.facebook.com/justia" title="Facebook" target="_blank" class="fb icon"><img src="https://lawyers.justia.com/s/facebook.svg" width="50" height="50" alt="Facebook" data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Social Media - Facebook"></a>
 <a href="https://twitter.com/justiacom" title="Twitter" target="_blank" class="twitter icon"><img src="https://lawyers.justia.com/s/twitter.svg" width="50" height="50" alt="Twitter" data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Social Media - Twitter"></a>
 <a href="https://www.linkedin.com/company/justia" title="LinkedIn" target="_blank" class="linkedin icon"><img src="https://lawyers.justia.com/s/linkedin.svg" width="50" height="50" alt="LinkedIn" data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Social Media - LinkedIn"></a>
 <a href="https://lawyers.justia.com/organization/justia-inc-16302" title="Justia Lawyer Directory" class="justia icon"><img src="https://lawyers.justia.com/s/justia.svg" width="50" height="50" alt="Justia" data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Social Media - Justia Lawyer Directory"></a>
 <a href="https://www.youtube.com/justia" title="Justia Lawyer Directory" target="_blank" class="youtube icon"><img src="https://lawyers.justia.com/s/youtube.svg" width="50" height="50" alt="YouTube" data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Social Media - YouTube"></a>
 </span>
 <span class="credits"> &copy; 2023 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Copyright" href="https://www.justia.com/">Justia</a></span>
 </span>
 <span class="nav">
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Justia Connect" href="https://connect.justia.com/">Justia Connect</a>
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Legal Portal" href="https://www.justia.com/">Legal Portal</a>
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Company" href="https://company.justia.com/">Company</a>
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Help" href="https://support.justia.com/">Help</a>
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Terms of Service" href="https://www.justia.com/terms-of-service/">Terms of Service</a>
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Privacy Policy" href="https://www.justia.com/privacy-policy/">Privacy Policy</a>
 <a data-gtm-type="click" data-gtm-category="Navigation" data-gtm-action="Footer" data-gtm-label="Marketing Solutions" class="nowrap" href="https://www.justia.com/marketing/">Marketing Solutions</a>
 </span>
 </div>
 </div>
 </div>
 </div>
 <script src="https://justatic.com/v/20231003111152/shared/js/core.js"></script>
 <script async src="//pagead2.googlesyndication.com/pagead/js/adsbygoogle.js"></script>
 <script>
 (adsbygoogle = window.adsbygoogle || []).push({});
 </script>
 </body>
</html>