Related Experiment Videos
Specific monoclonal antibody against human trypsin
Carmen Acosta Bas1, Ileana B Baluja Conde, A I Brito Moreno
1Centro de Inmunoensayo, Apdo 6653, Ave 25 y 134, Cubanacán, Playa, Ciudad Habana, Cuba. iqmonoclo@cie.sld.cu
A monoclonal antibody (MAb) directed against human trypsin-1 has been produced by hybridization of myeloma cells with spleen cells of BALB/c immunized mice. Antibodies were screened by ultramicro enzymelinked immunosorbent assay (UMELISA). MAb was purified by affinity chromatography on protein A-Sepharose, and MAb had a high affinity for trypsin-1 with the affinity constant equal 1.79 x 10(9) L/mol. Specificity was studied by UMELISA using cross-reactant proteins; MAb gave a positive reaction with native trypsinogen-1 and with the same protein after reduction. Antibody appeared to be directed against sequential epitope. One-step purification is described. The method evolved the adsorption of the enzyme onto a Sepharose-MAb(3H9) affinity column. The collected fraction was characterized and is available for immunization of BALB/c mice and for the preparation of a standard for immunoenzymatic assay.
A monoclonal antibody (MAb) directed against human trypsin-1 has been produced by hybridization of myeloma cells with spleen cells of BALB/c immunized mice. Antibodies were screened by ultramicro enzymelinked immunosorbent assay (UMELISA). MAb was purified by affinity chromatography on protein A-Sepharose, and MAb had a high affinity for trypsin-1 with the affinity constant equal 1.79 x 10(9) L/mol. Specificity was studied by UMELISA using cross-reactant proteins; MAb gave a positive reaction with native trypsinogen-1 and with the same protein after reduction. Antibody appeared to be directed against sequential epitope. One-step purification is described. The method evolved the adsorption of the enzyme onto a Sepharose-MAb(3H9) affinity column. The collected fraction was characterized and is available for immunization of BALB/c mice and for the preparation of a standard for immunoenzymatic assay.