Determination of binding constants of antibodies fractionated according to their affinity to insulin
Karulin, A.I.; Dikov, M.M.; Egorov, A.M.
Biokhimiia 51(5): 746-750
1986
ISSN/ISBN: 0320-9725 PMID: 3518815 Document Number: 278136
The number of populations of antibodies differing in their affinity for insulin was determined in guinea pig antiserum. The antibody fractions differing in their affinity were obtained by affinity chromatography on an antigenic sorbent. Elution with a stepwise pH gradient from 4 to 2 differing in the number and form of the steps resulted in 10 fractions of antibodies obtained from individual animal sera. The binding constants of fractionated antibodies were determined by a bioluminescent immunocofactor method. With a change in elution pH from 4 to 2, the equilibrious binding constant of antibodies increased from 10(6) to 5 X 10(9) M-1.