Preparation of a novel monoclonal antibody specific for WIG-1 and detection of its expression pattern in human esophageal carcinoma.
We have efficiently generated the first monoclonal antibody (MAb) against the human WIG-1 (wild-type p53-induced gene 1) protein, an apoptosis-related protein consisting of three zinc finger domains. Protein A affinity chromatography was performed to purify MAb from mice production ascites. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA). One MAb designated GW-3E4 (IgG1), effective in detecting the nuclear protein in human esophageal squamous cell carcinoma (ESCC) tissues and EC109 cell line, was characterized by ELISA and Western immunoblotting. Thus, it binds to native WIG-1 protein and should be useful in studies of WIG-1 protein function and expression. By Western immunoblotting analysis of 20 patients with ESCC using the MAb, we found that the expression of WIG-1 protein in tumor tissue was significantly higher than in incision margin. The results showed that WIG-1 might be a novel modifier in esophageal carcinogenesis, and the WIG-1 MAb should be useful in further study of the mechanism in WIG-1-related physiological reactions.
Department of Thoracic Surgery, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, PR China.
Ying-Bo
YB
Zou
Wei
W
Guo
Yao-Guang
YG
Jiang
Ru-Wen
RW
Wang
Yun-Ping
YP
Zhao
Zheng
Z
Ma
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism*; D000918:Antibody Specificity; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000378:metabolism* / Q000473:pathology; D002352:Carrier Proteins / Q000276:immunology* / Q000378:metabolism*; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D004938:Esophageal Neoplasms / Q000378:metabolism* / Q000473:pathology; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009687:Nuclear Proteins / Q000276:immunology* / Q000378:metabolism*; D016601:RNA-Binding Proteins; D014018:Tissue Distribution; D017186:Titrimetry
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; C436503:ZMAT3 protein, human