PittsburghV1, Istex, Corpus, bibRecord, 001650

Measurement of anti‐CD154 monoclonal antibody in primate sera by competitive inhibition ELISA

Identifieur interne : 001650 ( Istex/Corpus ); précédent : 001649; suivant : 001651

Measurement of anti‐CD154 monoclonal antibody in primate sera by competitive inhibition ELISA

Auteurs : Hao-Chih Tai ; Nathalie Campanile ; Mohamed Ezzelarab ; David K. C. Cooper ; Carol Phelps

Source :

Xenotransplantation [ 0908-665X ] ; 2006-11.

RBID : ISTEX:6235D251BC97E57BFDEF6B2B7656B44B113ECA63

English descriptors

Abstract

Abstract: Background: Anti‐human CD154 monoclonal antibody (mAb)‐based regimens have been demonstrated to prevent T cell‐dependent elicited antibody response in baboon recipients of pig hematopoietic progenitor cells, organs and islets. Monitoring of anti‐CD154 mAb in serum is important to ensure maintenance of adequate levels and for adjusting dosage of the anti‐CD154 mAb. We describe a method for measuring the level in primate sera.

Url:

https://api.istex.fr/document/6235D251BC97E57BFDEF6B2B7656B44B113ECA63/fulltext/pdf

DOI: 10.1111/j.1399-3089.2006.00345.x

Links to Exploration step

ISTEX:6235D251BC97E57BFDEF6B2B7656B44B113ECA63

Le document en format XML

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<profileDesc><abstract xml:lang="en" style="main"><p><hi rend="bold">Abstract: </hi>
<hi rend="bold"> Background: </hi>
 Anti‐human CD154 monoclonal antibody (mAb)‐based regimens have been demonstrated to prevent T cell‐dependent elicited antibody response in baboon recipients of pig hematopoietic progenitor cells, organs and islets. Monitoring of anti‐CD154 mAb in serum is important to ensure maintenance of adequate levels and for adjusting dosage of the anti‐CD154 mAb. We describe a method for measuring the level in primate sera.</p>
<p><hi rend="bold">Methods: </hi>
 The anti‐CD154 mAb level in primate serum was measured with a competitive inhibition enzyme linked immunosorbent assay in which the extent of inhibition of binding by anti‐CD154 mAb conjugated to horseradish peroxidase (anti‐CD154‐HRP) to soluble CD154 was used to determine the serum level. Briefly, a 96‐well maxisorb plate coated with soluble human CD154, and blocked with bovine serum albumin, was loaded with graded doses of anti‐CD154 mAb or primate sera containing anti‐CD154 mAb. Both were mixed with a known dosage of anti‐CD154‐HRP before loading. Bound anti‐CD154‐HRP was detected by color developed using 3,3′,5,5′ tetramethyl‐benzidine as substrate. Absorbance was measured in a Synergy<hi rend="superscript">TM</hi>
 HT Multi‐Detection Microplate Reader at a wavelength of 450 nm. Data analysis was carried out using BioTek's KC4<hi rend="superscript">TM</hi>
 Data Analysis Software. The standard curve was generated from the wells loaded with the mixture of anti‐CD154 mAb and anti‐CD154‐HRP.</p>
<p><hi rend="bold">Results and conclusions: </hi>
 The assay has been used successfully to measure anti‐CD154 mAb levels in the serum of both baboons and monkeys.</p>
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<titleGroup><title type="main" sort="XENOTRANSPLANTATION">Xenotransplantation</title>
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<publicationMeta level="part" position="11006"><doi origin="wiley">10.1111/xen.2006.13.issue-6</doi>
<numberingGroup><numbering type="journalVolume" number="13">13</numbering>
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<titleGroup><title type="tocHeading1">B<sc>rief</sc>
 C<sc>ommunication</sc>
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<numbering type="pageLast" number="570">570</numbering>
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<correspondenceTo>Address reprint requests to D. K. C. Cooper, MD, PhD, FRCS, Thomas E. Starzl Transplantation Institute, University of Pittsburgh Medical Center, Biomedical Science Tower; E1550A, 200 Lothrop Street, Pittsburgh, PA 15261, USA (E‐mail: <email>cooperdk@upmc.edu</email>
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<contentMeta><unparsedEditorialHistory>Received 2 June 2006; Accepted 14 July 2006</unparsedEditorialHistory>
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<titleGroup><title type="main">Measurement of anti‐CD154 monoclonal antibody in primate sera by competitive inhibition ELISA</title>
<title type="shortAuthors"><b>Tai et al.</b>
</title>
<title type="short"><b>Anti‐CD154 mAb levels</b>
</title>
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<creators><creator creatorRole="author" xml:id="cr1" affiliationRef="#a1"><personName><givenNames>Hao‐Chih</givenNames>
<familyName>Tai</familyName>
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<creator creatorRole="author" xml:id="cr2" affiliationRef="#a1"><personName><givenNames>Nathalie</givenNames>
<familyName>Campanile</familyName>
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<creator creatorRole="author" xml:id="cr3" affiliationRef="#a1"><personName><givenNames>Mohamed</givenNames>
<familyName>Ezzelarab</familyName>
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<creator creatorRole="author" xml:id="cr4" affiliationRef="#a1"><personName><givenNames>David K. C.</givenNames>
<familyName>Cooper</familyName>
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<creator creatorRole="author" xml:id="cr5" affiliationRef="#a2"><personName><givenNames>Carol</givenNames>
<familyName>Phelps</familyName>
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<affiliationGroup><affiliation xml:id="a1" countryCode="US"><unparsedAffiliation>The Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA, USA</unparsedAffiliation>
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<affiliation xml:id="a2" countryCode="US"><unparsedAffiliation>Revivicor, Inc., Blacksburg, VA, and Pittsburgh, PA, USA</unparsedAffiliation>
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<keywordGroup xml:lang="en"><keyword xml:id="k1">anti‐CD154 mAb</keyword>
<keyword xml:id="k2">costimulatory blockade</keyword>
<keyword xml:id="k3">enzyme linked immunosorbent assay</keyword>
<keyword xml:id="k4">competitive inhibition</keyword>
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<abstractGroup><abstract type="main" xml:lang="en"><!--
Tai H-C, Campanile N, Ezzelarab M, Cooper DKC, Phelps C. Measurement of anti-CD154 monoclonal antibody in primate sera by competitive inhibition ELISA.

Xenotransplantation 2006; 13: 566–570. © Blackwell Munksgaard, 2006
--><p><b>Abstract: </b>
<b> Background: </b>
 Anti‐human CD154 monoclonal antibody (mAb)‐based regimens have been demonstrated to prevent T cell‐dependent elicited antibody response in baboon recipients of pig hematopoietic progenitor cells, organs and islets. Monitoring of anti‐CD154 mAb in serum is important to ensure maintenance of adequate levels and for adjusting dosage of the anti‐CD154 mAb. We describe a method for measuring the level in primate sera.</p>
<p><b>Methods: </b>
 The anti‐CD154 mAb level in primate serum was measured with a competitive inhibition enzyme linked immunosorbent assay in which the extent of inhibition of binding by anti‐CD154 mAb conjugated to horseradish peroxidase (anti‐CD154‐HRP) to soluble CD154 was used to determine the serum level. Briefly, a 96‐well maxisorb plate coated with soluble human CD154, and blocked with bovine serum albumin, was loaded with graded doses of anti‐CD154 mAb or primate sera containing anti‐CD154 mAb. Both were mixed with a known dosage of anti‐CD154‐HRP before loading. Bound anti‐CD154‐HRP was detected by color developed using 3,3′,5,5′ tetramethyl‐benzidine as substrate. Absorbance was measured in a Synergy<sup>TM</sup>
 HT Multi‐Detection Microplate Reader at a wavelength of 450 nm. Data analysis was carried out using BioTek's KC4<sup>TM</sup>
 Data Analysis Software. The standard curve was generated from the wells loaded with the mixture of anti‐CD154 mAb and anti‐CD154‐HRP.</p>
<p><b>Results and conclusions: </b>
 The assay has been used successfully to measure anti‐CD154 mAb levels in the serum of both baboons and monkeys.</p>
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<name type="personal"><namePart type="given">Hao‐Chih</namePart>
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<abstract>Abstract:  Background:  Anti‐human CD154 monoclonal antibody (mAb)‐based regimens have been demonstrated to prevent T cell‐dependent elicited antibody response in baboon recipients of pig hematopoietic progenitor cells, organs and islets. Monitoring of anti‐CD154 mAb in serum is important to ensure maintenance of adequate levels and for adjusting dosage of the anti‐CD154 mAb. We describe a method for measuring the level in primate sera.</abstract>
<abstract>Methods:  The anti‐CD154 mAb level in primate serum was measured with a competitive inhibition enzyme linked immunosorbent assay in which the extent of inhibition of binding by anti‐CD154 mAb conjugated to horseradish peroxidase (anti‐CD154‐HRP) to soluble CD154 was used to determine the serum level. Briefly, a 96‐well maxisorb plate coated with soluble human CD154, and blocked with bovine serum albumin, was loaded with graded doses of anti‐CD154 mAb or primate sera containing anti‐CD154 mAb. Both were mixed with a known dosage of anti‐CD154‐HRP before loading. Bound anti‐CD154‐HRP was detected by color developed using 3,3′,5,5′ tetramethyl‐benzidine as substrate. Absorbance was measured in a SynergyTM HT Multi‐Detection Microplate Reader at a wavelength of 450 nm. Data analysis was carried out using BioTek's KC4TM Data Analysis Software. The standard curve was generated from the wells loaded with the mixture of anti‐CD154 mAb and anti‐CD154‐HRP.</abstract>
<abstract>Results and conclusions:  The assay has been used successfully to measure anti‐CD154 mAb levels in the serum of both baboons and monkeys.</abstract>
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<topic>costimulatory blockade</topic>
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Measurement of anti‐CD154 monoclonal antibody in primate sera by competitive inhibition ELISA

Measurement of anti‐CD154 monoclonal antibody in primate sera by competitive inhibition ELISA

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