Clinical Chemistry, Vol 31, 1654-1658, Copyright © 1985 by American Association for Clinical Chemistry

Monoclonal antibodies can precipitate low-density lipoprotein. I. Characterization and use in determining apolipoprotein B [published erratum appears in Clin Chem 1986 Feb;32(2):390]

S Marcovina, D France, RA Phillips and SJ Mao

We produced 20 mouse monoclonal antibodies against human plasma low- density lipoprotein (LDL). Individually they failed to precipitate LDL in agarose gel by the double-immunodiffusion technique; collectively they did, or as few as two combined monoclonal antibodies could do so. To mimic polyclonal antibodies in determination of apolipoprotein B (apo B) by radial immunodiffusion, a combination of four particular monoclonal antibodies (clones A, B, C, and D) was necessary. We characterized these four clones with respect to temperature dependency, affinity, total binding to 125I-labeled LDL, and specificity to the different species of apolipoprotein B. Two monoclonal antibodies (B and C) bound 100% of 125I-labeled LDL; clones A and D bound 80% and 87%, respectively. All four clones bound maximally to LDL at 4 degrees C. The affinity constants for clones A, B, C, and D were 0.6, 2.1, 3.8, and 2.3 X 10(9) L/mol, respectively. By the Western blotting technique, the four monoclonal antibodies all reacted with the species B-100 and B- 74 of apolipoprotein B, and to various degrees with B-48 and B-26. Radial immunodiffusion (chi) and direct enzyme-linked immunosorbent assay (y) with a mixture of the four monoclonal antibodies gave almost identical results for 70 patients: y = 0.921 chi-2.58; r = 0.933.

The following articles in journals at HighWire Press have cited this article:

				W. L. Chen, M. T. Huang, H. C. Liu, C. W. Li, and S. J. T. Mao Distinction Between Dry and Raw Milk Using Monoclonal Antibodies Prepared Against Dry Milk Proteins J Dairy Sci, August 1, 2004; 87(8): 2720 - 2729. [Abstract] [Full Text] [PDF]