Monoclonal antibody against human trypsin: production, characterization, and use for diagnosis.
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.
Acosta Bas
Carmen
C
Centro de Inmunoensayo, Apdo 6653, Ave 25 y 134, Cubanacán, Playa, Ciudad Habana, Cuba. iqmonoclo2@cie.sld.cu
Baluja Conde
Ileana B
IB
Brito Moreno
Adys I
AI
Rodríguez López
Maya Rosa
MR
Melchor
Antonio
A
Hernández
Liliana
L
Frómeta
Amarilys
A
Selles
María E
ME
Martínez
Luisa
L
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
D000911:Antibodies, Monoclonal; D014362:Trypsinogen; D014357:Trypsin