Defined Analyte-Enzyme Conjugates as Signal Generators in Immunoassays

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dc.contributor.author Paek, S. H. en_US
dc.contributor.author Bachas, L. G. en_US
dc.contributor.author Schramm, W. en_US
dc.date.accessioned 2006-04-10T15:48:27Z
dc.date.available 2006-04-10T15:48:27Z
dc.date.issued 1993-04 en_US
dc.identifier.citation Paek S. H., , Bachas L. G., , Schramm W., (1993/04)."Defined Analyte-Enzyme Conjugates as Signal Generators in Immunoassays." Analytical Biochemistry 210(1): 145-154. <http://hdl.handle.net/2027.42/30861> en_US
dc.identifier.uri http://www.sciencedirect.com/science/article/B6W9V-45R7D5F-57/2/db5f442a3a55adcac9585b0f752c049b en_US
dc.identifier.uri http://hdl.handle.net/2027.42/30861
dc.identifier.uri http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=8489010&dopt=citation en_US
dc.description.abstract We investigated the synthesis of progesterone-horseradish peroxidase (P-HRP) conjugates and products purified by affinity chromatography. The obtained preparations were characterized with an immobilized monoclonal antibody in solid-phase immunoassays. Three homogeneous P-HRP conjugates were isolated. Two preparations were identified to contain a single progesterone ligand on the enzyme molecule. A third preparation contained two progesterone ligands. We postulate that conjugation can occur at two different positions on the enzyme, and that the different microenvironment of the protein structure surrounding the ligand contributes to different binding constants of the conjugates with immunoglobulin. By comparing the effective binding constants derived from affinity chromatography and from Scatchard analysis, we have demonstrated that the divalent conjugate binds to antibody immobilized on planar surfaces only by a single attachment due to steric restriction. Dose-response curves for progesterone using the isolated P-HRP conjugates have been investigated and compared. en_US
dc.format.extent 999878 bytes
dc.format.extent 3118 bytes
dc.format.mimetype application/pdf
dc.format.mimetype text/plain
dc.language.iso en_US
dc.publisher Elsevier en_US
dc.title Defined Analyte-Enzyme Conjugates as Signal Generators in Immunoassays en_US
dc.type Article en_US
dc.rights.robots IndexNoFollow en_US
dc.subject.hlbsecondlevel Public Health en_US
dc.subject.hlbsecondlevel Chemistry en_US
dc.subject.hlbsecondlevel Chemical Engineering en_US
dc.subject.hlbsecondlevel Biological Chemistry en_US
dc.subject.hlbtoplevel Engineering en_US
dc.subject.hlbtoplevel Science en_US
dc.subject.hlbtoplevel Health Sciences en_US
dc.description.peerreviewed Peer Reviewed en_US
dc.contributor.affiliationum Reproductive Science and Bioengineering Program, University of Michigan, 300 N Ingalls, Ann Arbor, MI 48109, USA. en_US
dc.contributor.affiliationum Reproductive Science and Bioengineering Program, University of Michigan, 300 N Ingalls, Ann Arbor, MI 48109, USA; Bioquant Inc, Ann Arbor, MI 48105, USA. en_US
dc.contributor.affiliationother Department of Chemistry, University of Kentucky, Lexington, KY 40506, USA. en_US
dc.identifier.pmid 8489010 en_US
dc.description.bitstreamurl http://deepblue.lib.umich.edu/bitstream/2027.42/30861/1/0000524.pdf en_US
dc.identifier.doi http://dx.doi.org/10.1006/abio.1993.1165 en_US
dc.identifier.source Analytical Biochemistry en_US
dc.owningcollname Interdisciplinary and Peer-Reviewed
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