We used enzyme immunoassay to measure apolipoprotein B concentration in human plasma. Pure lipoprotein B was isolated from serum samples of fasting normolipidemic subjects by sequential preparative ultracentrifugation and coated to a polystyrene tube surface by adsorption. Human serum samples and rabbit antiserum to human apolipoprotein B were incubated with the solid-phase lipoprotein B. Soluble antigen competed with solid-phase antigen for binding to antibodies. After washing, peroxidase-labeled sheep antibodies against rabbit immunoglobulins were added, and after further washing the bound label was assayed. This provided a direct measurement of the soluble antigen. The best technical conditions for the assay were determined. The minimum detectable concentration was 1 microgram per assay. The enzyme immunoassay yielded values that compare favorably with those obtained by radial immunodiffusion (r = 0.84) and by rocket immunoelectrophoresis (r = 0.80). The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, ane non-use of radioisotopes.
%0 Journal Article
%1 citeulike:558743
%A Fruchart, J. C.
%A Desreumaux, C.
%A Dewailly, P.
%A Sezille, G.
%A Jaillard, J.
%A Carlier, Y.
%A Bout, D.
%A Capron, A.
%D 1978
%J Clin Chem
%K competition immunoassay elisa apob
%N 3
%P 455--459
%T Enzyme immunoassay for human apolipoprotein B, the major protein moiety in low-density- and very-low-density lipoproteins.
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=204430
%V 24
%X We used enzyme immunoassay to measure apolipoprotein B concentration in human plasma. Pure lipoprotein B was isolated from serum samples of fasting normolipidemic subjects by sequential preparative ultracentrifugation and coated to a polystyrene tube surface by adsorption. Human serum samples and rabbit antiserum to human apolipoprotein B were incubated with the solid-phase lipoprotein B. Soluble antigen competed with solid-phase antigen for binding to antibodies. After washing, peroxidase-labeled sheep antibodies against rabbit immunoglobulins were added, and after further washing the bound label was assayed. This provided a direct measurement of the soluble antigen. The best technical conditions for the assay were determined. The minimum detectable concentration was 1 microgram per assay. The enzyme immunoassay yielded values that compare favorably with those obtained by radial immunodiffusion (r = 0.84) and by rocket immunoelectrophoresis (r = 0.80). The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, ane non-use of radioisotopes.
@article{citeulike:558743,
abstract = {We used enzyme immunoassay to measure apolipoprotein B concentration in human plasma. Pure lipoprotein B was isolated from serum samples of fasting normolipidemic subjects by sequential preparative ultracentrifugation and coated to a polystyrene tube surface by adsorption. Human serum samples and rabbit antiserum to human apolipoprotein B were incubated with the solid-phase lipoprotein B. Soluble antigen competed with solid-phase antigen for binding to antibodies. After washing, peroxidase-labeled sheep antibodies against rabbit immunoglobulins were added, and after further washing the bound label was assayed. This provided a direct measurement of the soluble antigen. The best technical conditions for the assay were determined. The minimum detectable concentration was 1 microgram per assay. The enzyme immunoassay yielded values that compare favorably with those obtained by radial immunodiffusion (r = 0.84) and by rocket immunoelectrophoresis (r = 0.80). The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, ane non-use of radioisotopes.},
added-at = {2006-07-07T01:10:50.000+0200},
author = {Fruchart, J. C. and Desreumaux, C. and Dewailly, P. and Sezille, G. and Jaillard, J. and Carlier, Y. and Bout, D. and Capron, A.},
biburl = {https://www.bibsonomy.org/bibtex/28ae681e108fad050944c1b7b76dc1260/biblio24},
citeulike-article-id = {558743},
interhash = {ecdac005612f9cc1d6ba084746a4ad70},
intrahash = {8ae681e108fad050944c1b7b76dc1260},
issn = {0009-9147},
journal = {Clin Chem},
keywords = {competition immunoassay elisa apob},
month = {March},
number = 3,
pages = {455--459},
priority = {2},
timestamp = {2006-07-07T01:10:50.000+0200},
title = {Enzyme immunoassay for human apolipoprotein B, the major protein moiety in low-density- and very-low-density lipoproteins.},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=204430},
volume = 24,
year = 1978
}