| Structural determinants of cobrotoxin for binding to an enzyme-linked-immunoassay plate. | |
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MedLine Citation:
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PMID: 8756398 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In order to assess the manner in which the structural state of a protein affects the results of an ELISA, the antigenic reactivities of cobrotoxin and reduced and S-carboxymethylated (RCM-) cobrotoxin with anti-RCM-cobrotoxin antibodies were studied. The results of competitive enzyme-linked immunoassay showed that the affinity of RCM-cobrotoxin for anti-RCM-cobrotoxin antibodies was higher than that of cobrotoxin. However, the cobrotoxin-coated wells had a greater reactivity towards anti-RCM-cobrotoxin antibodies than against RCM-cobrotoxin-coated wells. The lower reactivity observed with RCM-cobrotoxin-coated plates could be improved by adding 0.01% glutaraldehyde during the coating procedure. Studies on the antigenic structures of RCM-cobrotoxin showed that it contained an immunodominant epitope at residues 22-38. Moreover, the N-terminal and C-terminal regions of RCM-cobrotoxin encompassed other antigenic determinants which exhibited low reactivities towards anti-RCM-cobrotoxin antibodies. After removal of the antibodies against residues 22-38 of cobrotoxin from anti-RCM-cobrotoxin antibodies by passage through an affinity column, the remaining antibodies exhibited a similar reactivity towards cobrotoxin and RCM-cobrotoxin. The antibodies against residues 22-38 retained a little reactivity with the RCM-cobrotoxin-coated wells. These results suggest that the structural determinants of cobrotoxin and RCM-cobrotoxin for binding to the microtitre plates differ. Unlike RCM-cobrotoxin, the loop II structure of cobrotoxin encompassing residues 22-38 is not exclusively involved in the binding of cobrotoxin to microtitre plates. |
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Authors:
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L S Chang; J Lin; C C Chang |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Biotechnology and applied biochemistry Volume: 24 ( Pt 1) ISSN: 0885-4513 ISO Abbreviation: Biotechnol. Appl. Biochem. Publication Date: 1996 Aug |
Date Detail:
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Created Date: 1996-09-20 Completed Date: 1996-09-20 Revised Date: 2007-11-15 |
Medline Journal Info:
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Nlm Unique ID: 8609465 Medline TA: Biotechnol Appl Biochem Country: ENGLAND |
Other Details:
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Languages: eng Pagination: 89-93 Citation Subset: IM |
Affiliation:
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Department of Biochemistry, Kaohsiung Medical College, Taiwan, Republic of China. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Antibodies Biotechnology Cobra Neurotoxin Proteins / chemistry*, immunology* Enzyme-Linked Immunosorbent Assay / methods* Epitopes / chemistry Methylation Molecular Structure Oxidation-Reduction Protein Folding |
| Chemical | |
Reg. No./Substance:
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0/Antibodies; 0/Cobra Neurotoxin Proteins; 0/Epitopes |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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