TY - JOUR
T1 - Monoclonal antibodies can precipitate low-density lipoprotein. I. Characterization and use in determining apolipoprotein B
AU - Marcovina, S.
AU - France, D.
AU - Phillips, R. A.
AU - Mao, S. J.T.
PY - 1985
Y1 - 1985
N2 - 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°C. The affinity constants for clones A, B, C, and D were 0.6, 2.1, 3.8, and 2.3 x 109 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 (x) and direct enzyme-linked immunosorbent assay (y) with a mixture of the four monoclonal antibodies gave almost identical results for 70 patients: y = 0.921x - 2.58; r = 0.933.
AB - 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°C. The affinity constants for clones A, B, C, and D were 0.6, 2.1, 3.8, and 2.3 x 109 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 (x) and direct enzyme-linked immunosorbent assay (y) with a mixture of the four monoclonal antibodies gave almost identical results for 70 patients: y = 0.921x - 2.58; r = 0.933.
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U2 - 10.1093/clinchem/31.10.1654
DO - 10.1093/clinchem/31.10.1654
M3 - Article
C2 - 3930091
AN - SCOPUS:0022204813
SN - 0009-9147
VL - 31
SP - 1654
EP - 1658
JO - Clinical Chemistry
JF - Clinical Chemistry
IS - 10
ER -