Studies on the immunological cross-reactivity of various pancreatic colipases. Isolation by immunoaffinity chromatography of a single form of procolipase from porcine pancreas.
Antibodies against porcine procolipase B were produced in rabbits. The antiserum was used to immunoinactivate various forms of native and trypsin-treated porcine colipase. Our results indicate that all forms of the porcine cofactor bind to anti-porcine procolipase B antibodies. Human colipase showed lower affinity for the antibodies than porcine colipase. No cross-reactivity was observed between pig and horse, cow, dog or chicken colipases. Immunological studies on porcine colipase, carried out in the presence of lipid, provided evidence that antibodies bind to colipase at or near the lipase binding site. The binding of antibodies to colipase is not affected by the adsorption of the cofactor at a lipid interface. Using a predictive method for identification of the antigenic determinants, it was found that, in pig colipase, regions at positions 42-48 and 70-74 might represent antigenic sites. In the horse protein, the peptide segment 42-48 was also recognized as a possible antigenic site. An immunoadsorbent gel column was prepared for a one-step isolation of porcine colipase. In contrast to purification methods described so far, immunoaffinity chromatography yielded only one form of the porcine cofactor when starting from a pancreatic extract. This protein preparation has structural, biochemical and immunochemical properties similar to that of porcine procolipase A previously isolated from pancreas in the presence of detergent.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D002846:Chromatography, Affinity; D003089:Colipases / Q000276:immunology / Q000302:isolation & purification*; D003429:Cross Reactions; D004792:Enzyme Precursors; D000939:Epitopes; D010179:Pancreas / Q000032:analysis*; D011498:Protein Precursors / Q000276:immunology / Q000302:isolation & purification*; D011506:Proteins / Q000302:isolation & purification*; D013552:Swine
D000906:Antibodies; D000936:Antigen-Antibody Complex; D003089:Colipases; D004792:Enzyme Precursors; D000939:Epitopes; D011498:Protein Precursors; D011506:Proteins; C037107:procolipase