Specific monoclonal antibody against human trypsin.
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.
Acosta Bas
Carmen
C
Centro de Inmunoensayo, Apdo 6653, Ave 25 y 134, Cubanacán, Playa, Ciudad Habana, Cuba. iqmonoclo@cie.sld.cu
Baluja Conde
Ileana B
IB
Brito Moreno
A I
AI
Rodriguez Lopez
Maya Rosa
MR
Melchor
Antonio
A
Hernandez
Liliana
L
Frómeta
Amarilys
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D000918:Antibody Specificity; D000939:Epitopes; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D014357:Trypsin / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D014357:Trypsin