Removal of blood group determinants from bovine erythrocyte membranes. 3. Action of proteolytic enzymes on intact cells

Abstract

SummaryBovine erythrocyte treatment with chymotrypsin, trypsin, pronase, papain, or ficin eliminated or weakened the reactivity of 18 of the 47 blood group factors which were examined. Thirteen of the affected factors were from the B system, and one each was from the C, FV, L, M, and R'S' systems. Variation attributable to pheno‐group (allele) or genotype influences was observed in the effects upon six of the factors. Ficin‐treated V/V, but not F/V or F/F, cells were rapidly lysed by normal rabbit serum (complement control). Absorptions with pronase‐treated V positive cells indicated that essentially all V antigenicity was removed. However, immunizations with pronase‐treated V positive cells elicited V antibody production in one of two recipient cows. The numbers of antigens removed by different enzymes did not appear to be closely related to the amount of protein removed.

Apparent destruction of bovine blood group determinants by chymotrypsin

“…Haemolytic tests of washed, enzyme-treated red cells verified that digestions had occurred and that results were as reported by Hines et al (1976). I n addition to the expected results, one serendipitous observation was made.…”

“…dures of Hines et al (1976). In some experiments 0.05 % or 0.025 % (w/v) chymotrypsin concentrations with two hours incubation were substituted for the original 0.1 % (w/v) concentration with 30 minutes of incubation.…”

“…The haemolytic test procedures have been described (Hines et al, 1976). Haemolytic inhibition tests were similarly performed in-plastic microtiter plates, with 0.025 ml quantities of product and antisera being incubated for 30 minutes at 24 "C prior to the addition of red cells.…”