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Journal of Lipid Research, Vol 24, 1639-1645, Copyright © 1983 by Lipid Research, Inc.
DL Koritnik and LL Rudel
A sensitive and specific enzyme-linked immunosorbent assay (ELISA) for
nonhuman primate serum apolipoprotein A-I (apoA-I) is described. The assay
is a noncompetitive, sandwich ELISA in which polystyrene microtiter plates
were used with purified, monospecific goat anti- monkey apoA-I antibodies
adsorbed on the wells. The serum samples were added to the coated wells,
incubated, and after washing, antibodies conjugated to horseradish
peroxidase were added. After further washing, the bound label was assayed.
A heat treatment step, 52 degrees C for 3 hr, was used to maximize the
apoA-I immunoreactive sites in diluted serum. Serum samples extracted with
chloroform-methanol, delipidated with tetramethylurea, or denatured by
heating gave essentially equivalent results. The working range of the
apoA-I standards was 0.5 to 5 ng and parallel responses were observed for
apoA-I in serum, in isolated HDL, and in buffer as a purified apoprotein.
Recovery of apoA- I added to serum was quantitative (106 +/- 3%). The
intra- and interassay coefficients of variation were 6.2 and 6.9%,
respectively. The enzyme immunoassay yielded values that compared favorably
with those obtained by radial immunodiffusion (r = 0.84). ApoA-I
concentration in African green monkey serum was highly correlated with the
HDL cholesterol concentration (r = 0.86). It is concluded that this ELISA
is an accurate and precise method for determination of apoA-I
concentrations in primate serum.
ARTICLES
Measurement of apolipoprotein A-I concentration in nonhuman primate serum by enzyme-linked immunosorbent assay (ELISA)
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