Preparation and characterization of a novel monoclonal antibody specific to human EMILIN-5 protein.
We aimed to establish the first monoclonal antibody (MAb) against the human EMILIN-5 protein (elastin microfibril interface located protein-5) and to investigate its distribution in normal human esophageal tissues and esophageal carcinomas. The bacterially expressed 6 His-EMILIN-5 fusion protein was induced and purified. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using either purified 6x His-EMILIN-5 fusion proteins or purified 6x His-ZNRD1 fusion protein as a control. The EMILIN-5 protein-specific MAb was further identified by Western immunoblot analysis. The expression of EMILIN-5 was investigated in 63 cases of esophageal cancer specimens and 60 cases of normal esophageal specimens using immunohistochemical analysis. The expression of 6x His-EMILIN-5 fusion proteins was successfully induced. One MAb, H7 (IgG1), effective in detecting the recombinant and the cellular protein, was characterized. H7 bound to native EMILIN-5 protein and should be useful in studies of EMILIN-5 protein function and expression. EMILIN-5 staining was found positive in 37 cases of esophageal cancer tissues (59%) and 7 cases of normal esophageal tissues (12%). The higher-grade frequency of expression of EMILIN-5 in normal esophageal tissues was significantly lower (p < 0.01) than that in tumor tissues. We concluded that EMILIN-5 might play important roles in carcinogenesis of esophageal cancer.
Cao
Jun
J
Department of Orthodontics, Qindu Stomatology College Affiliated to the Fourth Military Medical University, Xi'an, Shaanxi Province, China. zhanganuo@126.com
D000368:Aged; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry*; D000954:Antigens, Surface / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D045744:Cell Line, Tumor; D004938:Esophageal Neoplasms / Q000737:chemistry / Q000209:etiology / Q000378:metabolism; D005260:Female; D006801:Humans; D006825:Hybridomas; D008297:Male; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000737:chemistry / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; C089826:EMILIN3 protein, human; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; C030340:elastin microfibril interface located protein