Enzyme-linked immunosorbent assay for human plasma apolipoprotein B.
A noncompetitive enzyme-linked immunosorbent assay (ELISA) has been developed for measuring total plasma apolipoprotein (apo) B using affinity purified polyclonal and monoclonal antibodies. Microtiter plates from different manufacturers were tested with regard to their IgG binding characteristics; only one plate yielded consistent coefficients of variation of less than 5%. The optimal plasma dilution in this assay was 1:3000. IgG anti-apoB antisera conjugated to alkaline phosphatase was used as a second antibody. p-Nitrophenyl phosphate was utilized as substrate for color development, and the absorbance (410 nm) was read utilizing an ELISA reader interfaced with a microcomputer for data processing. Plasma apoB levels in plasma have been determined in 1115 male and female participants in the Framingham Offspring Study. Mean (+/- SD) plasma concentrations were 89 +/- 28 mg/dl. Significant age and sex related differences in apoB levels were noted.
Ordovas
J M
JM
USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA.
Peterson
J P
JP
Santaniello
P
P
Cohn
J S
JS
Wilson
P W
PW
Schaefer
E J
EJ
D000328:Adult; D000367:Age Factors; D000368:Aged; D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B / Q000097:blood*; D002784:Cholesterol / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D012016:Reference Values; D012737:Sex Factors; D013048:Specimen Handling
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D002784:Cholesterol