Direct double antibody sandwich immunoassay of mucin M1 epitopes in human mucus secreting pancreatic cell lines.
A direct sandwich enzyme immunoassay using two monoclonal antibodies was developed in order to quantify mucin M1 antigens produced by two pancreatic adenocarcinoma cell lines: CAPAN-1 and CFPAC-1. As a solid phase, the wells of a microtiter plate were coated with a first monoclonal antibody, 1-13 M1 and the biotinylated monoclonal conjugate 9-13 M1 was used as the second antibody. The assay was optimized with streptavidin-peroxidase. The detection limit of the assay is 1.6 ng/ml. This ELISA is highly specific, sensitive, reproducible and quickly performed. It will permit the comparison of mucin exocytosis by the two cell lines in response to secretagogue agents and may help in the study of the pathogenesis of mucus hypersecretion such as cystic fibrosis.
Montserrat
C
C
Groupe de Recherche sur les Glandes Exocrines, Faculté de Médecine, Marseille, France.
Hollande
E
E
Guy-Crotte
O
O
Figarella
C
C
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001426:Bacterial Proteins; D001710:Biotin; D002460:Cell Line; D003550:Cystic Fibrosis / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D009077:Mucins / Q000737:chemistry* / Q000276:immunology; D009093:Mucus / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D010179:Pancreas / Q000378:metabolism*; D010190:Pancreatic Neoplasms / Q000378:metabolism; D011506:Proteins / Q000032:analysis; D019809:Streptavidin; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D000939:Epitopes; D009077:Mucins; D011506:Proteins; D001710:Biotin; D019809:Streptavidin