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Influence of iodothyronine conjugates of bovine serum albumin and horseradish peroxidase on enzyme immunosorbent assay of thyroid hormones.
MedLine Citation:
PMID:  24295178     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
Enzyme-linked immunosorbent assays (ELISA's) reported for thyroxine (T4) and 3,5,3'-triiodothyronine (T3), involved coupling of the haptens through (i) carboxylic group to carrier protein for producing antibodies and (ii) amino group to detection labels. To improve the titer and specificity of antibodies, immunogens were prepared by coupling of carboxyl group to bovine serum albumin (BSA) either directly or through adipic acid dihydrazide (ADH), after protecting amino group through acetylation of T4 and T3. Direct coupling resulted in the incorporation of 40-50 moles of T4 and T3 per BSA molecule and helped in improving immunogenic response and specificity, especially of T4. High epitope density of immunogens evoked better antibody response, since attachement of ADH as spacer, introduced 18-27 moles of haptens into carrier protein and had less effect on antibody development, with T3 being exception. Detection labels were prepared by coupling horseradish peroxidase (HRP) to amino group of thyroid hormones directly and after preparing their methyl esters, which provided sensitive displacement curves in combination with the antibodies developed against N-acetylated-T4 and T3. Unlike methyl esters, T4-HRP and T3-HRP showed higher sensitivity and seemed to be related to the affinity of the labels for binding the antibody.
Authors:
G Lakshmi Kumari; Sachin Kumar; Satish Gupta; Anuradha Saini; Sudesh K Sharma; Navneet Kaur
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of immunoassay & immunochemistry     Volume:  35     ISSN:  1532-4230     ISO Abbreviation:  J Immunoassay Immunochem     Publication Date:  2014 Apr 
Date Detail:
Created Date:  2013-12-03     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100963688     Medline TA:  J Immunoassay Immunochem     Country:  England    
Other Details:
Languages:  eng     Pagination:  139-56     Citation Subset:  IM    
Affiliation:
a Applied Biotech Engineering Center , Ambala College of Engineering and Applied Research , Ambala , Haryana , India.
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