Clinical Chemistry
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Clinical Chemistry 41: 1150-1158, 1995;
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Clinical Chemistry, Vol 41, 1150-1158, Copyright © 1995 by American Association for Clinical Chemistry

Characterization of a monoclonal antibody (HB-22) and development of an ELISA for human apolipoprotein A-I

TC Rothwell, VS Kamanna, FY Jin, E Koren, T Foley and ML Kashyap
Hycor Biomedical Inc., Garden Grove, CA 92641, USA.

We describe the production and characterization of a high-affinity monoclonal antibody, HB-22, for apolipoprotein (apo) A-I, a major protein of human high-density lipoproteins (HDL). Including Tween 20 in the reaction mixture increased the binding capacity of HB-22 to apo A- I. HB-22 showed monospecific reactivity with HDL or apo A-I, displaying no cross-reactivity with apo A-II, intermediate-, low-, or very-low- density lipoproteins. Immunoaffinity columns with HB-22 (in the absence of Tween 20) showed an immunosorbent capacity of 80 micrograms of apo A- I per milligram of antibody. The immunosorbent capacity of HB-22 for apo A-I was similar in plasma samples from normolipidemic, hypercholesterolemic, or hypertriglyceridemic patients. Comparative binding studies demonstrated that compared with other available monoclonal apo A-I antibodies, HB-22 had the greatest apparent affinity for binding to HDL. A competitive ELISA developed by utilizing HB-22 could detect as little as 20 ng of apo A-I in the reaction mixture. The intra- and interassay CVs of the ELISA were 5.4% and 9.5%, respectively.


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Copyright © 1995 by the American Association for Clinical Chemistry.