Subtilopeptidase A isoenzyme system. Interaction with serum components and its importance for quantitative immunoelectrophoresis.
A method was developed which involved electroimmunoassay and crossed immunoelectrophoresis of subtilopeptidase A (EC 3.4.21.14). Initial trials with unfractionated antiserum were not successful and interaction of the enzyme with non-immunoglobulin serum components were shown to be the cause of the failures. Quantitative immunoelectrophoresis was possible when purified immunoglobulins were used. A pH of 6.5 (lower than the usual pH 8.6) was necessary to obtain a proper baseline definition. Subtilopeptidase A was confirmed as a multiple isoenzyme system. Qualitative inter-batch variations were detected. Di-isopropyl phosphorofluoridate inhibition altered the electrophoretic pattern, but no loss of antigenic determinants was observed.
D000818:Animals; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007123:Immunoelectrophoresis, Two-Dimensional; D007136:Immunoglobulins / Q000302:isolation & purification; D007527:Isoenzymes / Q000032:analysis*; D007531:Isoflurophate / Q000494:pharmacology; D011817:Rabbits; D013381:Subtilisins / Q000032:analysis* / Q000037:antagonists & inhibitors
D007136:Immunoglobulins; D007527:Isoenzymes; D007531:Isoflurophate; D013381:Subtilisins