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<meta name="robots" content="INDEX,FOLLOW,NOARCHIVE" /><meta name="citation_inbook_title" content="Molecular Imaging and Contrast Agent Database (MICAD) [Internet]" /><meta name="citation_title" content="99mTc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody" /><meta name="citation_publisher" content="National Center for Biotechnology Information (US)" /><meta name="citation_date" content="2008/01/28" /><meta name="citation_author" content="Kenneth T. Cheng" /><meta name="citation_pmid" content="20641631" /><meta name="citation_fulltext_html_url" content="https://www.ncbi.nlm.nih.gov/books/NBK23432/" /><link rel="schema.DC" href="http://purl.org/DC/elements/1.0/" /><meta name="DC.Title" content="99mTc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody" /><meta name="DC.Type" content="Text" /><meta name="DC.Publisher" content="National Center for Biotechnology Information (US)" /><meta name="DC.Contributor" content="Kenneth T. Cheng" /><meta name="DC.Date" content="2008/01/28" /><meta name="DC.Identifier" content="https://www.ncbi.nlm.nih.gov/books/NBK23432/" /><meta name="description" content="99mTc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody (99mTc-HYNIC-CC49 sc(Fv)2 MAb), which is formed by the conjugation of 99mTc with a bioengineered antitumor-associated glycoprotein 72 (TAG-72) antibody construct, has been developed for single-photon emission computed tomography (SPECT) imaging of cancers that express TAG-72 (1). 99mTc is a gamma emitter with a half-life (t½) of 6.02 h." /><meta name="og:title" content="99mTc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody" /><meta name="og:type" content="book" /><meta name="og:description" content="99mTc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody (99mTc-HYNIC-CC49 sc(Fv)2 MAb), which is formed by the conjugation of 99mTc with a bioengineered antitumor-associated glycoprotein 72 (TAG-72) antibody construct, has been developed for single-photon emission computed tomography (SPECT) imaging of cancers that express TAG-72 (1). 99mTc is a gamma emitter with a half-life (t½) of 6.02 h." /><meta name="og:url" content="https://www.ncbi.nlm.nih.gov/books/NBK23432/" /><meta name="og:site_name" content="NCBI Bookshelf" /><meta name="og:image" content="https://www.ncbi.nlm.nih.gov/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" /><meta name="twitter:card" content="summary" /><meta name="twitter:site" content="@ncbibooks" /><meta name="bk-non-canon-loc" content="/books/n/micad/CC49ScFv2Tc99m/" /><link rel="canonical" href="https://www.ncbi.nlm.nih.gov/books/NBK23432/" /><link rel="stylesheet" href="/corehtml/pmc/css/figpopup.css" type="text/css" media="screen" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books.min.css" type="text/css" /><link rel="stylesheet" href="/corehtml/pmc/css/bookshelf/2.26/css/books_print.min.css" type="text/css" media="print" /><style type="text/css">p a.figpopup{display:inline !important} .bk_tt {font-family: monospace} .first-line-outdent .bk_ref {display: inline} .body-content h2, .body-content .h2 {border-bottom: 1px solid #97B0C8} .body-content h2.inline {border-bottom: none} a.page-toc-label , .jig-ncbismoothscroll a {text-decoration:none;border:0 !important} .temp-labeled-list .graphic {display:inline-block !important} .temp-labeled-list img{width:100%}</style><script type="text/javascript" src="/corehtml/pmc/js/jquery.hoverIntent.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/common.min.js?_=3.18"> </script><script type="text/javascript" src="/corehtml/pmc/js/large-obj-scrollbars.min.js"> </script><script type="text/javascript">window.name="mainwindow";</script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/book-toc.min.js"> </script><script type="text/javascript" src="/corehtml/pmc/js/bookshelf/2.26/books.min.js"> </script><meta name="book-collection" content="NONE" />
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<div class="pre-content"><div><div class="bk_prnt"><p class="small">NCBI Bookshelf. A service of the National Library of Medicine, National Institutes of Health.</p><p>Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. </p></div><div class="iconblock clearfix whole_rhythm no_top_margin bk_noprnt"><a class="img_link icnblk_img" title="Table of Contents Page" href="/books/n/micad/"><img class="source-thumb" src="/corehtml/pmc/pmcgifs/bookshelf/thumbs/th-micad-lrg.png" alt="Cover of Molecular Imaging and Contrast Agent Database (MICAD)" height="100px" width="80px" /></a><div class="icnblk_cntnt eight_col"><h2>Molecular Imaging and Contrast Agent Database (MICAD) [Internet].</h2><a data-jig="ncbitoggler" href="#__NBK23432_dtls__">Show details</a><div style="display:none" class="ui-widget" id="__NBK23432_dtls__"><div>Bethesda (MD): <a href="https://www.ncbi.nlm.nih.gov/" ref="pagearea=page-banner&amp;targetsite=external&amp;targetcat=link&amp;targettype=publisher">National Center for Biotechnology Information (US)</a>; 2004-2013.</div></div><div class="half_rhythm"><ul class="inline_list"><li style="margin-right:1em"><a class="bk_cntns" href="/books/n/micad/">Contents</a></li></ul></div><div class="bk_noprnt"><form method="get" action="/books/n/micad/" id="bk_srch"><div class="bk_search"><label for="bk_term" class="offscreen_noflow">Search term</label><input type="text" title="Search this book" id="bk_term" name="term" value="" data-jig="ncbiclearbutton" /> <input type="submit" class="jig-ncbibutton" value="Search this book" submit="false" style="padding: 0.1em 0.4em;" /></div></form></div></div><div class="icnblk_cntnt two_col"><div class="pagination bk_noprnt"><a class="active page_link prev" href="/books/n/micad/Annexin-HYNIC-99mTc/" title="Previous page in this title">&lt; Prev</a><a class="active page_link next" href="/books/n/micad/CC49ScFv4Tc99m/" title="Next page in this title">Next &gt;</a></div></div></div></div></div>
<div class="main-content lit-style" itemscope="itemscope" itemtype="http://schema.org/CreativeWork"><div class="meta-content fm-sec"><h1 id="_NBK23432_"><span class="title" itemprop="name"><sup>99m</sup>Tc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody </span></h1><div itemprop="alternativeHeadline" class="subtitle whole_rhythm"><sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb</div><p class="contrib-group"><span itemprop="author">Kenneth T. Cheng</span>, PhD.</p><a data-jig="ncbitoggler" href="#__NBK23432_ai__" style="border:0;text-decoration:none">Author Information and Affiliations</a><div style="display:none" class="ui-widget" id="__NBK23432_ai__"><div class="contrib half_rhythm"><span itemprop="author">Kenneth T. Cheng</span>, PhD<div class="affiliation small">
National Center for Biotechnology Information, NLM, NIH, Bethesda, MD,
<span class="before-email-separator"></span><span class="email-label">Email: </span><a href="mailto:dev@null" data-email="vog.hin.mln.ibcn@dacim" class="oemail">vog.hin.mln.ibcn@dacim</a>
</div></div></div><p class="small">Created: <span itemprop="datePublished">June 19, 2007</span>; Last Update: <span itemprop="dateModified">January 28, 2008</span>.</p></div><div class="jig-ncbiinpagenav body-content whole_rhythm" data-jigconfig="allHeadingLevels: ['h2'],smoothScroll: false" itemprop="text"><div id="CC49ScFv2Tc99m.T1" class="table"><p class="large-table-link" style="display:none"><span class="right"><a href="/books/NBK23432/table/CC49ScFv2Tc99m.T1/?report=objectonly" target="object">View in own window</a></span></p><div class="large_tbl" id="__CC49ScFv2Tc99m.T1_lrgtbl__"><table><tbody><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Chemical name:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><sup>99m</sup>Tc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody</td><td rowspan="9" colspan="1" style="text-align:left;vertical-align:middle;"></td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Abbreviated name:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Synonym:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><sup>99m</sup>Tc-CC49 sc(Fv)<sub>2</sub> MAb, <sup>99m</sup>Tc-CC49 MAb</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Agent Category:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Divalent single-chain Fv monoclonal antibody (sc(Fv)<sub>2</sub> MAb</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Target:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">TAG-72</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Target Category:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Antibody to antigen binding</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Method of detection:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Single-photon emission computed tomography (SPECT), planar gamma imaging</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Source of signal/ contrast:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;"><sup>99m</sup>Tc</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Activation:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">No</td></tr><tr><td rowspan="1" colspan="1" style="text-align:right;vertical-align:top;">
<b>Studies:</b>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">
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<img alt="Checkbox" src="/corehtml/pmc/css/bookshelf/2.26/img/studies.checkbox.png" /> Rodents
</div></li></ul>
</td><td rowspan="1" colspan="1" style="text-align:left;vertical-align:top;">Click on <a href="/entrez/viewer.fcgi?db=protein&#x00026;id=144964805" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">protein</a>, <a href="/entrez/viewer.fcgi?db=nuccore&#x00026;id=21689921" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">nucleotide</a> (RefSeq), and <a href="/sites/entrez?Db=gene&#x00026;Cmd=ShowDetailView&#x00026;TermToSearch=182875&#x00026;ordinalpos=1&#x00026;itool=EntrezSystem2.PEntrez.Gene.Gene_ResultsPanel.Gene_RVDocSum" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">gene</a> for more information about TAG-72</td></tr></tbody></table></div></div><div id="CC49ScFv2Tc99m.Background"><h2 id="_CC49ScFv2Tc99m_Background_">Background</h2><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22SUBSTANCENAME%22%5BSubstance%20Name%5D" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p><sup>99m</sup>Tc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody (<sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb), which is formed by the conjugation of <sup>99m</sup>Tc with a bioengineered anti&#x02013;tumor-associated glycoprotein 72 (TAG-72) antibody construct, has been developed for single-photon emission computed tomography (SPECT) imaging of cancers that express TAG-72 (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>). <sup>99m</sup>Tc is a gamma emitter with a half-life (<i>t</i><sub>&#x000bd;</sub>) of 6.02 h.</p><p>The TAG-72 antigen was isolated from the LS-174T human colon cancer xenograft as a high molecular weight glycoprotein (molecular mass of 10<sup>6</sup> Da) with mucin-like characteristics (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.2">2-5</a>). It is expressed on a variety of human adenocarcinomas such as pancreatic, breast, colorectal, prostate, endometrial, and ovarian cancers. This antigen has also been shown to be shed into the serum of cancer patients (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.6">6</a>). The murine MAb B72.3 against TAG-72 was initially generated by immunization of mice with a membrane-enriched fraction of a human breast carcinoma (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.7">7</a>). With use of affinity-purified TAG-72 from LS-174T as an immunogen, CC49 and other anti&#x02013;TAG-2 MAbs with higher affinity constants (<i>K</i><sub>a</sub>) have been produced and characterized (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1-3</a>, <a class="bk_pop" href="#CC49ScFv2Tc99m.REF.7">7</a>).</p><p>Radiolabeled MAbs have been developed for both the diagnosis and treatment of tumors (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.8">8</a>). Radiolabeled B72.3 and CC49 have shown excellent tumor localization capabilities with potential diagnostic and therapeutic applications in the clinical setting (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.9">9</a>, <a class="bk_pop" href="#CC49ScFv2Tc99m.REF.10">10</a>). Because of their relatively large size, radiolabeled intact MAbs tend to have unfavorable imaging kinetics, poor tumor penetration, and high potential for human anti-mouse antibody response (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>, <a class="bk_pop" href="#CC49ScFv2Tc99m.REF.11">11-13</a>). One approach to minimize these problems is reducing intact antibodies to antibody fragments such as F(ab&#x02019;)<sub>2</sub> and Fab&#x02019; (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.14">14</a>). Another approach is the development of genetic engineering methods to obtain single-chain Fv constructs (scFv) and multivalent scFv constructs (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>, <a class="bk_pop" href="#CC49ScFv2Tc99m.REF.15">15</a>, <a class="bk_pop" href="#CC49ScFv2Tc99m.REF.16">16</a>). These scFv constructs contain the variable regions of the light chain (V<sub>L</sub>) and heavy chain (V<sub>H</sub>) connected by a flexible linker. Colcher et al. (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.17">17</a>) constructed the monomeric CC49 scFv MAb (~27 kDa), which selectively recognizes a unique sialyl-Tn epitope of TAG-72. The radioiodinated CC49 scFv appeared to clear rapidly from the blood with good tumor penetration (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.16">16</a>, <a class="bk_pop" href="#CC49ScFv2Tc99m.REF.18">18</a>). To further improve the imaging kinetics, Pavlinkova et al. (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.18">18</a>) constructed the high-affinity dimer CC49 sc(Fv)<sub>2</sub> (~60 kDa). The radioiodinated CC49 sc(Fv)<sub>2</sub> showed good stability and increased avidity <i>in vivo</i> compared with the radioiodinated CC49 scFv construct.</p><p>With direct or indirect labeling, MAbs can be labeled with <sup>99m</sup>Tc, a gamma emitter with ideal SPECT imaging properties. Direct labeling involves reduction of <sup>99m</sup>Tc-pertechnetate and nonspecific binding of the reduced <sup>99m</sup>Tc to donor atoms, namely thiol, amide, amino, and carboxylate (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.19">19</a>). Indirect labeling uses a bifunctional chelating agent, which can be more binding site&#x02013;specific on the MAb molecule. Goel et al. (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>) used hydrazinonicotinamide (HYNIC) as a bifunctional coupling agent to label CC49 sc(Fv)<sub>2</sub> with <sup>99m</sup>Tc. The <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb showed good tumor targeting and <i>in vivo</i> biodistribution properties.</p></div><div id="CC49ScFv2Tc99m.Synthesis"><h2 id="_CC49ScFv2Tc99m_Synthesis_">Synthesis</h2><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20synthesis" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p>Goel et al. (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>) reported the construction and radiolabeling of the <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb. The CC49 scFv (V<sub>L</sub>-linker-V<sub>H</sub>) was derived from the murine MAb CC49 cloned in the yeast expression vector pPICZ&#x003b1;A and constructed with the 205C linker. The bacterial scFv construct was used as the template DNA for the expression of the scFv in competent <i>P. pastoris</i> KM71 cells. The construction of the divalent sc(Fv)<sub>2</sub> (V<sub>L</sub>-linker-V<sub>H</sub>-linker-V<sub>L</sub>-linker-V<sub>H</sub>-His<sub>6</sub>) was performed as described by Goel et al. (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.20">20</a>) using the 205C linker in a <i>P. pastoris</i> expression system, and the final construct was purified from the secreted medium with immobilized metal affinity chromatography. The preparation was shown to be &#x0003e;95% pure by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Size-exclusion high-performance liquid chromatography (HPLC) showed the molecular mass to be 60 kDa. Competitive solid-phase competition enzyme-linked immunosorbent assay (ELISA) with bovine submaxillary gland mucin (BSM) confirmed the immunoreactivity of the divalent construct. To radiolabel the agent, the hydrazino-modification of CC49 sc(Fv)<sub>2</sub> was achieved by reacting the construct with the <i>N</i>-hydroxysuccinimide ester of succinimidyl-6-hydrazinonicotinate hydrochloride (SHNH) at a molar ratio of 10:1 in sodium phosphate buffer (pH 7.8). It was estimated that there were ~2.3 SHNH groups per sc(Fv)<sub>2</sub>. In the radiolabeling procedure, sodium <sup>99m</sup>Tc-pertechnetate, tricine, and stannous chloride were first mixed, then SHNH-derivatized CC49 sc(Fv)<sub>2</sub> was added, and the reaction mixture was incubated at room temperature for 45 min. The final <sup>99m</sup>Tc-HYNIC-CC49 sc(FV)<sub>2</sub> MAb was purified on a Sephadex G-25 column. The specific activity was 74&#x02013;111 MBq/mg (2&#x02013;3 mCi/mg) or 4.44&#x02013;6.66 MBq/nmol (0.12&#x02013;0.18 mCi/nmol) on the basis of the estimated 60-kDa molecular weight) with a radiochemical purity &#x02265;95% (HPLC analysis). On SDS-PAGE, &#x0003e;90% of the total radioactivity was associated with the protein band of 58 kDa.</p></div><div id="CC49ScFv2Tc99m.In_Vitro_Studies_Tes"><h2 id="_CC49ScFv2Tc99m_In_Vitro_Studies_Tes_"><i>In Vitro</i> Studies: Testing in Cells and Tissues</h2><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20in%20vitro" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p>The immunoreactivity of <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb by solid-phase ELISA with immobilized BSM was 85&#x02013;95% with a nonspecific binding of 0.8&#x02013;1.5% (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>).</p><p><i>In vitro</i> stability studies of <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb were conducted by incubating the radiolabeled MAb construct in 1% bovine serum albumin (BSA) or 1% mouse serum at 37&#x000ba;C for 24 h (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>). By HPLC analysis, &#x0003c;20% loss of <sup>99m</sup>Tc label was detected in 1% BSA. In the 1% mouse serum, ~20% and 10% of the <sup>99m</sup>Tc were associated with low molecular weight proteins (&#x0003c;50 kDa) and high molecular weight proteins (&#x0003e;130 kDa), respectively. This latter protein fraction suggested the possibility of aggregation of the radiolabel with serum proteins.</p><p>Using the Scatchard plot and surface plasmon resonance technique to measure the real-time interactions, Goel et al. (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.21">21</a>) reported the <i>K</i><sub>a</sub> of unlabeled CC49 sc(Fv)<sub>2</sub> to be 2.75 &#x000d7; 10<sup>7</sup> M&#x02212;<sup>1</sup> in binding to the immobilized BSM. In comparison, the <i>K</i><sub>a</sub> for the intact CC49 MAb construct was 1.14 &#x000d7; 10<sup>8</sup> M&#x02212;<sup>1</sup>.</p></div><div id="CC49ScFv2Tc99m.Animal_Studies"><h2 id="_CC49ScFv2Tc99m_Animal_Studies_">Animal Studies</h2><div id="CC49ScFv2Tc99m.Rodents"><h3>Rodents</h3><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20rodentia" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p>Biodistribution studies of <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb were performed in nude mice bearing LS-147T s.c. human colon carcinomas (~250&#x02013;300 mm<sup>3</sup>) (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>). Each mouse received 0.37 MBq (10 &#x003bc;Ci) of <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> mAb by i.v. administration. The blood elimination <i>t</i><sub>&#x000bd;</sub> was 144 min, and the whole-body clearance <i>t</i><sub>&#x000bd;</sub> was 184 &#x000b1; 19 min (<i>n</i> = 3). The radioactivity levels (<i>n</i> = 3 &#x000d7; 2) in percentage injected dose per gram (% ID/g) of the tumors were 9.9 &#x000b1; 0.7 (0.5 h), 10.9 &#x000b1; 0.8 (1 h), 12.6 &#x000b1; 0.4 (4 h), 7.2 &#x000b1; 0.7 (8 h), 2.3 &#x000b1; 0.1 (16 h), and 1.2 &#x000b1; 0.0 (24 h). At 16 h, the tumor/blood ratio was 4.6:1. At 0.5 h, the radioactivity levels (% ID/g) of major organs were 13.9 &#x000b1; 1.1 (blood), 18.9 &#x000b1; 1.5 (liver), 13.4 &#x000b1; 1.2 (spleen), 33.9 &#x000b1; 1.8 (kidneys), and 5.4 &#x000b1; 0.8 (lungs). At 4 h, these levels changed to 4.3 &#x000b1; 0.2 (blood), 14.8 &#x000b1; 1.3 (liver), 9.7 &#x000b1; 0.7 (spleen), 27.3 &#x000b1; 1.4 (kidneys), and 1.3 &#x000b1; 0.0 (lungs). By 24 h, these levels declined to 0.1 &#x000b1; 0.0 (blood), 1.2 &#x000b1; 0.0 (liver), 1.0 &#x000b1; 0.0 (spleen), 2.4 &#x000b1; 0.3 (kidneys), and 0.0 &#x000b1; 0.0 (lungs). Macroautoradiography studies performed in mice at 6 h and 16 h after radioactivity administration confirmed a high degree of tumor localization and negligible retention in the blood and normal organs (<a class="bk_pop" href="#CC49ScFv2Tc99m.REF.1">1</a>). The exceptions were the liver and pancreas, but the tumor radioactivity levels were still two-fold higher at both 6 and 16 h in these two organs. The tumor remained positive at 16 h after injection. The study suggested that <sup>99m</sup>Tc-HYNIC-CC49 sc(Fv)<sub>2</sub> MAb probably underwent hepatobiliary excretion.</p></div><div id="CC49ScFv2Tc99m.Other_NonPrimate_Mam"><h3>Other Non-Primate Mammals</h3><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22%20SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20%28dog%20OR%20rabbit%20OR%20pig%20OR%20sheep%29" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div><div id="CC49ScFv2Tc99m.NonHuman_Primates"><h3>Non-Human Primates</h3><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20%28primate%20NOT%20human%29" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div></div><div id="CC49ScFv2Tc99m.Human_Studies"><h2 id="_CC49ScFv2Tc99m_Human_Studies_">Human Studies</h2><p>[<a href="/entrez/query.fcgi?cmd=PureSearch&#x00026;db=pubmed&#x00026;details_term=%22SUBSTANCENAME%22%5BSubstance%20Name%5D%20AND%20human" ref="pagearea=body&amp;targetsite=external&amp;targetcat=link&amp;targettype=uri">PubMed</a>]</p><p>No publication is currently available.</p></div><div id="CC49ScFv2Tc99m.references"><h2 id="_CC49ScFv2Tc99m_references_">References</h2><dl class="temp-labeled-list"><dt>1.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.1">Goel A. , Baranowska-Kortylewicz J. , Hinrichs S.H. , Wisecarver J. , Pavlinkova G. , Augustine S. , Colcher D. , Booth B.J. , Batra S.K. 99mTc-labeled divalent and tetravalent CC49 single-chain Fv's: novel imaging agents for rapid in vivo localization of human colon carcinoma. <span><span class="ref-journal">J Nucl Med. </span>2001;<span class="ref-vol">
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</span>(10):151927.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11585867" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 11585867</span></a>]</div></dd><dt>2.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.2">Muraro R. , Kuroki M. , Wunderlich D. , Poole D.J. , Colcher D. , Thor A. , Greiner J.W. , Simpson J.F. , Molinolo A. , Noguchi P. et al. Generation and characterization of B72.3 second generation monoclonal antibodies reactive with the tumor-associated glycoprotein 72 antigen. <span><span class="ref-journal">Cancer Res. </span>1988;<span class="ref-vol">
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</span>(16):458896.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3396010" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3396010</span></a>]</div></dd><dt>3.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.3">Johnson V.G. , Schlom J. , Paterson A.J. , Bennett J. , Magnani J.L. , Colcher D. Analysis of a human tumor-associated glycoprotein (TAG-72) identified by monoclonal antibody B72.3. <span><span class="ref-journal">Cancer Res. </span>1986;<span class="ref-vol">
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</span>(2):8507.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3940648" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3940648</span></a>]</div></dd><dt>4.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.4">Katari R.S. , Fernsten P.D. , Schlom J. Characterization of the shed form of the human tumor-associated glycoprotein (TAG-72) from serous effusions of patients with different types of carcinomas. <span><span class="ref-journal">Cancer Res. </span>1990;<span class="ref-vol">
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</span>(16):488590.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/2379152" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 2379152</span></a>]</div></dd><dt>5.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.5">Xiao J. , Horst S. , Hinkle G. , Cao X. , Kocak E. , Fang J. , Young D. , Khazaeli M. , Agnese D. , Sun D. , Martin E. Pharmacokinetics and clinical evaluation of 125I-radiolabeled humanized CC49 monoclonal antibody (HuCC49deltaC(H)2) in recurrent and metastatic colorectal cancer patients. <span><span class="ref-journal">Cancer Biother Radiopharm. </span>2005;<span class="ref-vol">
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</span>(1):1626.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/15778575" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 15778575</span></a>]</div></dd><dt>6.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.6">Paterson A.J. , Schlom J. , Sears H.F. , Bennett J. , Colcher D. A radioimmunoassay for the detection of a human tumor-associated glycoprotein (TAG-72) using monoclonal antibody B72.3. <span><span class="ref-journal">Int J Cancer. </span>1986;<span class="ref-vol">
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</span>(5):65966.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3699929" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3699929</span></a>]</div></dd><dt>7.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.7">Colcher D. , Hand P.H. , Nuti M. , Schlom J. A spectrum of monoclonal antibodies reactive with human mammary tumor cells. <span><span class="ref-journal">Proc Natl Acad Sci U S A. </span>1981;<span class="ref-vol">
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</span>(5):3199203.</span> [<a href="/pmc/articles/PMC319528/" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pmc">PMC free article<span class="bk_prnt">: PMC319528</span></a>] [<a href="https://pubmed.ncbi.nlm.nih.gov/6789331" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 6789331</span></a>]</div></dd><dt>8.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.8">Kowalsky R.J. , Falen S.W. <span><span class="ref-journal">and Radiopharmaceuticals in nuclear pharmacy and nuclear medicine, American Pharmacists Association: Washington, D.C. p. 733-752. </span>2004</span></div></dd><dt>9.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.9">Colcher D. , Minelli M.F. , Roselli M. , Muraro R. , Simpson-Milenic D. , Schlom J. Radioimmunolocalization of human carcinoma xenografts with B72.3 second generation monoclonal antibodies. <span><span class="ref-journal">Cancer Res. </span>1988;<span class="ref-vol">
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</span>(16):4597603.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3396011" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3396011</span></a>]</div></dd><dt>10.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.10">Colcher D. , Esteban J. , Carrasquillo J.A. , Sugarbaker P. , Reynolds J.C. , Bryant G. , Larson S.M. , Schlom J. Complementation of intracavitary and intravenous administration of a monoclonal antibody (B72.3) in patients with carcinoma. <span><span class="ref-journal">Cancer Res. </span>1987;<span class="ref-vol">
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</span>(15):421824.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3607761" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3607761</span></a>]</div></dd><dt>11.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.11">Britton K.E. The development of new radiopharmaceuticals. <span><span class="ref-journal">Eur J Nucl Med. </span>1990;<span class="ref-vol">
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</span>(4-6):37385.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/2190837" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 2190837</span></a>]</div></dd><dt>12.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.12">Jain R.K. Transport of molecules across tumor vasculature. <span><span class="ref-journal">Cancer Metastasis Rev. </span>1987;<span class="ref-vol">
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</span>(3):497501.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/7008935" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 7008935</span></a>]</div></dd><dt>14.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.14">Behr T. , Becker W. , Hannappel E. , Goldenberg D.M. , Wolf F. <span class="ref-title">Targeting of liver metastases of colorectal cancer with IgG, F(ab')2, and Fab' anti-carcinoembryonic antigen antibodies labeled with 99mTc: the role of metabolism and kinetics </span><span class="ref-journal">Cancer Res</span> 1995<strong>55</strong>Suppl235777s5785s. [<a href="https://pubmed.ncbi.nlm.nih.gov/7493346" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 7493346</span></a>]</div></dd><dt>15.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.15">Bird R.E. , Hardman K.D. , Jacobson J.W. , Johnson S. , Kaufman B.M. , Lee S.M. , Lee T. , Pope S.H. , Riordan G.S. , Whitlow M. Single-chain antigen-binding proteins. <span><span class="ref-journal">Science. </span>1988;<span class="ref-vol">
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</span>(4877):4236.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3140379" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3140379</span></a>]</div></dd><dt>16.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.16">Colcher D. , Bird R. , Roselli M. , Hardman K.D. , Johnson S. , Pope S. , Dodd S.W. , Pantoliano M.W. , Milenic D.E. , Schlom J. In vivo tumor targeting of a recombinant single-chain antigen-binding protein. <span><span class="ref-journal">J Natl Cancer Inst. </span>1990;<span class="ref-vol">
<strong>82</strong>
</span>(14):11917.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/2362290" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 2362290</span></a>]</div></dd><dt>17.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.17">Colcher D. , Goel A. , Pavlinkova G. , Beresford G. , Booth B. , Batra S.K. Effects of genetic engineering on the pharmacokinetics of antibodies. <span><span class="ref-journal">Q J Nucl Med. </span>1999;<span class="ref-vol">
<strong>43</strong>
</span>(2):1329.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10429508" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10429508</span></a>]</div></dd><dt>18.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.18">Pavlinkova G. , Beresford G.W. , Booth B.J. , Batra S.K. , Colcher D. Pharmacokinetics and biodistribution of engineered single-chain antibody constructs of MAb CC49 in colon carcinoma xenografts. <span><span class="ref-journal">J Nucl Med. </span>1999;<span class="ref-vol">
<strong>40</strong>
</span>(9):153646.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10492377" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10492377</span></a>]</div></dd><dt>19.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.19">Fritzberg A.R. , Berninger R.W. , Hadley S.W. , Wester D.W. Approaches to radiolabeling of antibodies for diagnosis and therapy of cancer. <span><span class="ref-journal">Pharm Res. </span>1988;<span class="ref-vol">
<strong>5</strong>
</span>(6):32534.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/3072555" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 3072555</span></a>]</div></dd><dt>20.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.20">Goel A. , Beresford G.W. , Colcher D. , Pavlinkova G. , Booth B.J. , Baranowska-Kortylewicz J. , Batra S.K. Divalent forms of CC49 single-chain antibody constructs in Pichia pastoris: expression, purification, and characterization. <span><span class="ref-journal">J Biochem (Tokyo). </span>2000;<span class="ref-vol">
<strong>127</strong>
</span>(5):82936.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/10788792" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 10788792</span></a>]</div></dd><dt>21.</dt><dd><div class="bk_ref" id="CC49ScFv2Tc99m.REF.21">Goel A. , Colcher D. , Baranowska-Kortylewicz J. , Augustine S. , Booth B.J. , Pavlinkova G. , Batra S.K. Genetically engineered tetravalent single-chain Fv of the pancarcinoma monoclonal antibody CC49: improved biodistribution and potential for therapeutic application. <span><span class="ref-journal">Cancer Res. </span>2000;<span class="ref-vol">
<strong>60</strong>
</span>(24):696471.</span> [<a href="https://pubmed.ncbi.nlm.nih.gov/11156397" ref="pagearea=cite-ref&amp;targetsite=entrez&amp;targetcat=link&amp;targettype=pubmed">PubMed<span class="bk_prnt">: 11156397</span></a>]</div></dd></dl></div><div id="bk_toc_contnr"></div></div></div>
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<div xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Views</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="PDF_download" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="/books/NBK23432/?report=reader">PubReader</a></li><li><a href="/books/NBK23432/?report=printable">Print View</a></li><li><a data-jig="ncbidialog" href="#_ncbi_dlg_citbx_NBK23432" data-jigconfig="width:400,modal:true">Cite this Page</a><div id="_ncbi_dlg_citbx_NBK23432" style="display:none" title="Cite this Page"><div class="bk_tt">Cheng KT. 99mTc-Hydrazinonicotinamide-anti-TAG-72 CC49 divalent single-chain Fv monoclonal antibody. 2007 Jun 19 [Updated 2008 Jan 28]. In: Molecular Imaging and Contrast Agent Database (MICAD) [Internet]. Bethesda (MD): National Center for Biotechnology Information (US); 2004-2013. <span class="bk_cite_avail"></span></div></div></li><li><a href="/books/NBK23432/pdf/Bookshelf_NBK23432.pdf">PDF version of this page</a> (144K)</li><li><a href="/books/n/micad/toc/bin/micad.csv">MICAD summary (CSV file)</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>In this page</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="page-toc" id="Shutter"></a></div><div class="portlet_content"><ul xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" class="simple-list"><li><a href="#CC49ScFv2Tc99m.Background" ref="log$=inpage&amp;link_id=inpage">Background</a></li><li><a href="#CC49ScFv2Tc99m.Synthesis" ref="log$=inpage&amp;link_id=inpage">Synthesis</a></li><li><a href="#CC49ScFv2Tc99m.In_Vitro_Studies_Tes" ref="log$=inpage&amp;link_id=inpage"><i>In Vitro</i> Studies: Testing in Cells and Tissues</a></li><li><a href="#CC49ScFv2Tc99m.Animal_Studies" ref="log$=inpage&amp;link_id=inpage">Animal Studies</a></li><li><a href="#CC49ScFv2Tc99m.Human_Studies" ref="log$=inpage&amp;link_id=inpage">Human Studies</a></li><li><a href="#CC49ScFv2Tc99m.references" ref="log$=inpage&amp;link_id=inpage">References</a></li></ul></div></div><div class="portlet"><div class="portlet_head"><div class="portlet_title"><h3><span>Search MICAD</span></h3></div><a name="Shutter" sid="1" href="#" class="portlet_shutter" title="Show/hide content" remembercollapsed="true" pgsec_name="source-application" id="Shutter"></a></div><div class="portlet_content"><form xmlns:np="http://ncbi.gov/portal/XSLT/namespace" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" name="frmSearch" method="get" action="/books/NBK5330/" id="frmSearch"><script type="text/javascript" src="/corehtml/pmc//js/bookshelf/micad.js">/**/</script><label class="offscreen_noflow" for="txtfield">Search term</label><input id="txtfield" type="text" name="f1_term" size="22" onKeyPress="KeyPress('micad',event,'/books/NBK5330/','')" /><button name="f1_search" type="submit">Go</button><button onclick="this.form.reset();" type="reset">Clear</button><p><b>Limit my Search:</b></p><div class="clearfix"><label for="detection">Method of detection:</label><div class="right"><select name="detection" id="detection" style="width:200px"><option value="" selected="selected">Any</option><option value="(MRI OR &quot;Magnetic resonance imaging&quot; OR MRS)">MRI</option><option value="Multimodal">Multimodal imaging</option><option value="Optical">Optical imaging</option><option value="PET">PET</option><option value="Photoacoustic">Photoacoustic imaging</option><option value="(SPECT OR planar)">SPECT</option><option value="Ultrasound">Ultrasound</option><option value="(x-ray OR ct)">X-ray, CT</option></select></div></div><div class="clearfix"><label for="signal">Source of signal/contrast:</label><div class="right"><select name="signal" id="signal" style="width:200px"><option value="" selected="selected">Any</option><optgroup label="MRI agents"><option value="(Copper OR Cu)">Copper</option><option value="(Europium OR Eu3+)">Europium</option><option value="(Fluorine OR 19F)">Fluorine</option><option value="(Gadolinium OR Gd3+)">Gadolinium</option><option value="&quot;Hyperpolarized 13C&quot;">Hyperpolarized 13C</option><option value="&quot;Iron oxide&quot;">Iron oxide</option><option value="&quot;Nitroxide radicals&quot;">Nitroxide radicals</option><option value="(Oxygen OR 17O)">Oxygen</option><option value="Thulium">Thulium</option></optgroup><optgroup label="Multimodal agents"><option value="((Gadolinium OR Gd3+) AND Optical)">Gadolinium and optical</option><option value="((Gadolinium OR Gd3+) AND (Gold OR Au))">Gadolinium and Gold</option><option value="(&quot;Iron oxide&quot; AND (64Cu OR 124I OR 111In))">Iron oxide and
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