A specific and sensitive Sandwich enzyme-linked immunosorbent assay (ELISA) for the quantitative determination of apolipoprotein C-III, a major apolipoprotein of human very low density lipoproteins is described. The assay is non-competitive and it uses the same affinity isolated sheep antibody both for coating the wells and as conjugate with alkaline phosphatase. Total serum apo C-III was determined in a normal population of 24 men and 21 women. The difference was not statistically significant. In both sexes apo C-III concentration correlated positively with the serum triglyceride levels. In patients with hyperlipoproteinemia, apo C-III levels were increased.
References
1.
OsborneJCJr, BrewerH B. The plasma lipo proteins. In: AnfinsenCB, EdsallJT, RichardsFM, Eds. Advances in Protein Chemistry, Vol. 31. New York: Academic Press, 1979; 253–337.
AvogaroP. CazzolatoG, BittoloG Bon. QuinciG B. Are apolipoproteins better discriminators than lipids for atherosclerosis?Lancet1979; I: 901–3.
4.
RiesenWF, MordasiniR. SalzmannCApoproteins and lipids as discriminators of severity of coronary heart disease. Atherosclerosis1980; 37: 157–62.
5.
LaRosaJC, LevyRI. HerbertPNA specific apoprotein activator for lipoprotein lipase. Biochem Biophys Res Commun1970; 41: 57–62.
6.
HavelRJ, FieldingCJ, OlivereronaTCo-factor activity of protein components of human very low density lipoprotein in the hydrolysis of triglycerides by lipoprotein lipase from different sources. Biochemistry1973; 12: 1828–33.
7.
BreckenridgeWC, LittleJA. SteinerGHypertriglyceridemia associated with deficiency of apolipoprotein C-II. New Engl J Med1978; 298: 1265–73.
8.
BrownWV, BaginskyM L. Inhibition of lipoprotein lipase by an apoprotein of human very low density lipoprotein. Biochem Biophys Res Commun1972; 46: 375–82.
9.
KraussRM, HerbertPN, LevyRI, FredricksonD S. Further observations on the activation and inhibition of lipoprotein lipase by apolipoproteins. Circulation Res1973; 33: 403–11.
10.
ShelbourneF, HanksJ, MeyersW, QuarfordtS. Effect of apoproteins on hepatic uptake of triglyceride emulsions in the rat. J Clin Invest1980; 65: 652–8.
11.
WindierE, ChaoY, HavelR J. Regulation of the hepatic uptake of triglyceride-rich lipoproteins in the rat. J Biol Chem1980: 255: 8303–7.
12.
BursteinM, ScholnickHR, MorfinR. Rapid method for the isolation of lipoproteins from human serum by precipitation with polyanions. J Lipid Res1970; 11: 583–95.
13.
ScanuAM, EdelsteinC. Solubility in aqueous solutions of ethanol of the small molecular weight peptides of the serum very low density and high density lipoproteins: relevance to the recovery problem during delipidation of scrum lipoproteins. Anal Biochem1971; 44: 576–88.
MarcelYL, BergsethM, NestruckA C. Preparative isoelectric focussing of apolipoproteins C and E from human very low density lipoproteins. Biochim Biophys Acta1979; 573: 175–83.
16.
RiesenWF, MordasiniRC, MiddelhoffG W. Quantitation of the two major apoproteins of human high density lipoproteins by solid phase radioimmunoassay. FEBS Letters1978; 91: 35–9.
17.
EngvallE, PerlmannP. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry1971; 8: 871–4.
18.
KearneyJF, RadbruckA, LicsegangB, RajewskyF. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol1979; 123: 1548–50.
19.
SullivanMJ, GnemmiE, MorrisEA simple method for the preparation of enzyme-antibody conjugates. FEBS Letters1978; 95: 311–5.
20.
SchonfeldG, GeorgePK, MillerJApolipoprotein C-II and C-III levels in hyperlipo-protcinacmia. Metabolism1979; 28: 1001–10.
21.
KashyapML, SrivastavaLS, HyndBAQuantitation of human apolipoprotein C-III and its subspecies by radioimmunoassay and analytical isoelectric focusing: abnormal plasma triglyceride-rich lipoprotein apolipoprotein C-III subspecies concentration in hypertriglyceridemia. J Lipid Res1981; 22: 800–10.
22.
CurryMD, McConathyWJ, FesmireJD, AlaupovicP. Quantitative determination of human apolipoprotein C-III by electro-immunoassay. Biochim Biophys Acta1980; 617: 503–13.
23.
BuryJ, De KeersgieterW. RossenauM. Immunonephelometric quantitation of apolipoprotein C-III in plasma. Clin Chim Acta1985; 145: 249–58.
24.
HolmquistL.Quantitation of human serum very low density apolipoproteins C-I, C-II. C-III. and E by enzyme immunoassay. J Immunol Meth1980; 34: 243–51.
25.
ParsyD, ClaveyV, FievetC, KoraI, DuriezPQuantification of apolipoprotein C-III in serum by non-competitive immunoenzymometric assay. Clin Chem1985; 31: 1632–5.
26.
StarrJI, JunDJ, MakoMECirculating high density lipoprotein (HDL): Measurement of an apolipoprotein component by radioimmunoassay. J Clin Invest1976; 52: 819.
27.
KaneJP, SataT, HamiltonRL, HavelR J. Apoprotein composition of very low density lipoproteins of human serum. J Clin Invest1975; 56: 1622–34.
28.
BradleyDD, WingredJ, PetittiDBHigh-density lipoproteins, cholesterol and oral contraceptives. New Engl J Med1978; 299: 17–20.
29.
CheungMC, AlbersJ J. Measurement of apolipoprotein A-I and A-II levels in men and women by immunoassay. J Clin Invest1977; 60: 43–50.
30.
TikkanenMJ, NikkiläE A. Menopausal oestrogen therapy, serum lipoproteins and ischemic heart disease. Lancet1981; 1: 1319.
31.
WarnickGR, MayfieldC, AlbersJJ, HazzardW R. Gel isoelectric focusing method for specific diagnosis of familial hyperlipoproteinaemia type, 3. Clin Chem1979; 25: 279–84.
32.
PolzE, KostnerG M. The binding of B2-glycoprotein-I to human serum lipoproteins. Distribution among density fractions. FEBS Letters1979; 102: 183–6.
