title: Monoclonal antibody against human trypsin: production, characterization, and use for diagnosis.
abstract: A monoclonal antibody (MAb) directed against human immunoaffinity purified trypsinogen has been produced by hybridization of myeloma cells with spleen cells of BALB/c immunized mice. Antibodies were screened by ultramicro-enzyme-linked immunosorbent assay (UMELISA). The MAb was purified by affinity chromatography on protein A-sepharose, and MAb had a high affinity for trypsinogen with the affinity constant equal 2.06 x 10(9) L/mol. Specificity was studied by UMELISA using cross-reactant proteins; MAb gave a positive reaction with native trypsinogen-1 but did not react with the same protein after reduction. The antibody seem to be directed against conformational epitope. The MAb obtained was characterized immunologically and used to develop UMELISA for detection Trypsin. This monoclonal assay enabled the detection of 2.8 ng/mL.
authors.affiliation: Centro de Inmunoensayo, Apdo 6653, Ave 25 y 134, Cubanacán, Playa, Ciudad Habana, Cuba. iqmonoclo2@cie.sld.cu
authors.forename: Carmen
authors.initials: C
authors.lastname: Acosta Bas
authors.forename: Ileana B
authors.initials: IB
authors.lastname: Baluja Conde
authors.forename: Adys I
authors.initials: AI
authors.lastname: Brito Moreno
authors.forename: Maya Rosa
authors.initials: MR
authors.lastname: Rodríguez López
authors.forename: Antonio
authors.initials: A
authors.lastname: Melchor
authors.forename: Liliana
authors.initials: L
authors.lastname: Hernández
authors.forename: Amarilys
authors.initials: A
authors.lastname: Frómeta
authors.forename: María E
authors.initials: ME
authors.lastname: Selles
authors.forename: Luisa
authors.initials: L
authors.lastname: Martínez
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D003550:Cystic Fibrosis / Q000175:diagnosis; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007231:Infant, Newborn; D051379:Mice; D008807:Mice, Inbred BALB C; D015997:Neonatal Screening; D014357:Trypsin / Q000032:analysis / Q000097:blood / Q000276:immunology*; D014362:Trypsinogen / Q000032:analysis / Q000097:blood / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D014362:Trypsinogen; D014357:Trypsin