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<article dtd-version="3.0" article-type="Invited Methods Article - JoVE Produced Video">
  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher">jove</journal-id>
      <journal-id journal-id-type="email">editorial-office@jove.com</journal-id>
      <journal-title-group>
        <journal-title>Journal of Visualized Experiments</journal-title>
      </journal-title-group>
      <issn pub-type="ppub">1940-087X</issn>
      <issn pub-type="epub"></issn>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id">JoVE58830</article-id>
      <article-id pub-id-type="manuscript">JoVE58830</article-id>
      <article-categories>
        <subj-group subj-group-type="Article Type">
          <subject>Invited Methods Article - JoVE Produced Video</subject>
        </subj-group>
        <subj-group subj-group-type="Category">
          <subject>JoVE Bioengineering</subject>
        </subj-group>
        <subj-group subj-group-type="Classification">
          <subject>Analytical, Diagnostic and Therapeutic Techniques and Equipment</subject>
          <subject>Diagnosis</subject>
          <subject>Therapeutics</subject>
          <subject>Investigative Techniques</subject>
          <subject>Equipment and Supplies</subject>
          <subject>Life sciences</subject>
          <subject>Life Sciences (General)</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Detection of Endotoxin in Nano-formulations using Limulus Amoebocyte Lysate (LAL) assays</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <role content-type="1" />
          <name>
            <surname>Neun</surname>
            <given-names>Barry  W</given-names>
          </name>
          <email>neunb@mail.nih.gov</email>
          <xref ref-type="aff" rid="aff1" />
        </contrib>
        <contrib contrib-type="author" corresp="yes">
          <role content-type="2" />
          <name>
            <surname>Dobrovolskaia</surname>
            <given-names>Marina</given-names>
            <prefix>Dr.</prefix>
          </name>
          <email>marina@mail.nih.gov</email>
          <xref ref-type="aff" rid="aff2" />
        </contrib>
        <contrib contrib-type="editor">
          <name>
            <surname>Navani</surname>
            <given-names>Dipesh</given-names>
          </name>
          <degrees>Ph.D.</degrees>
          <email>dipesh.n@jove.com</email>
          <xref ref-type="aff" rid="aff3" />
        </contrib>
        <aff id="aff1">
          <institution>Frederick National Laboratory for Cancer Research</institution>
          <addr-line content-type="addrline1">1050 Boyles St</addr-line>
          <country>UNITED STATES</country>
          <addr-line content-type="city">Frederick</addr-line>
          <addr-line content-type="state">MD</addr-line>
          <addr-line content-type="zipcode">21701</addr-line>
        </aff>
        <aff id="aff2">
          <institution>Frederick National Laboratory for Cancer Research</institution>
          <institution content-type="dept">CRTP</institution>
          <addr-line content-type="addrline1">1050 Boyles St</addr-line>
          <country>UNITED STATES</country>
          <addr-line content-type="city">Frederick</addr-line>
          <addr-line content-type="state">MD</addr-line>
          <addr-line content-type="zipcode">21702</addr-line>
        </aff>
        <aff id="aff3">
          <institution>JoVE</institution>
          <addr-line>1 Alewife Center</addr-line>
          <addr-line>Suite 200</addr-line>
          <addr-line>Cambridge, MA  02140</addr-line>
          <country>UNITED STATES</country>
        </aff>
      </contrib-group>
      <history>
        <date date-type="received">
          <day>26</day>
          <month>07</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>17</day>
          <month>09</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>14</day>
          <month>09</month>
          <year>2018</year>
        </date>
        <date date-type="accepted">
          <day>19</day>
          <month>09</month>
          <year>2018</year>
        </date>
      </history>
      <abstract>
        <p>When present in pharmaceutical products, Gram-negative bacterial cell wall component endotoxin (often also called lipopolysaccharide) can cause inflammation, fever, hypo- or hypertension, and, in extreme cases, lead to tissue and organ damage which may become fatal. The amounts of endotoxin in pharmaceutical products, therefore, are strictly regulated. Among the methods available for endotoxin detection and quantification, Limulus Amoebocyte Lysate (LAL) assay is commonly used worldwide. While any pharmaceutical product can interfere with the LAL assay, nanoformulations represent a particular challenge due to their complexity. The purpose of this paper is to provide a practical guide to researchers inexperienced in estimating endotoxin in engineered nanomaterials and nanoparticle-formulated drugs. Herein, we describe practical recommendations for performing three LAL formats including turbidity, chromogenic and gel-clot assays. These assays can be used to determine endotoxin contamination in nanotechnology-based drug products, vaccines, and adjuvants</p>
      </abstract>
      <kwd-group>
        <kwd>endotoxin, pyrogen, nanomaterials, immunotoxicity, nanomedicine</kwd>
      </kwd-group>
      <funding-group />
      <counts>
        <fig-count count="0" />
      </counts>
      <custom-meta-group>
        <custom-meta>
          <meta-name>Please indicate whether this article will be Standard Access or Open Access. </meta-name>
          <meta-value>Standard Access (US$2,400);</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Please indicate the &lt;b&gt; city, state/province, and country&lt;/b&gt; where this article will be &lt;b&gt;filmed&lt;/b&gt;. Please do not use abbreviations.</meta-name>
          <meta-value>Frederick, MD</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Black and White Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Color Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Acquisitions Editor</meta-name>
          <meta-value>Nandita Singh</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Assistant who entered lead</meta-name>
          <meta-value>Jennifer Carroll</meta-value>
        </custom-meta>
      </custom-meta-group>
    </article-meta>
  </front>
  <body>
    <table-wrap>
      <label>Materials</label>
      <graphic xlink:href="JoVE_Materials.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Figure</label>
      <graphic xlink:href="Picture1.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Table 1</label>
      <graphic xlink:href="Picture1.jpg" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table 1</label>
      <graphic xlink:href="Table 1.xlsx" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
  </body>
</article>