title: Monoclonal antibodies directed against VP7 protein of human group B rotavirus.
abstract: The aim of this study was to prepare and identify a monoclonal antibody that binds the viral proteins 7 (VP7 protein) of human group B rotavirus (GBRV) and to describe its immunologic characterization. Human group B rotavirus vp7 gene was successfully ligated into pGEX-KG vector and transformed into Escherichia coli TOP10 cells. The glutathione S-transferases (GST)-fusion protein GST-VP7 was induced by Isopropyl β-D-1-thiogalactopyranoside (IPTG) and immediately purified to immunize BALB/c mice. Splenocytes were then prepared from the immunized mouse and fused with SP2/0 myeloma cell line. In the end we obtained one positive hybridoma cell line stably secreting monoclonal antibody against GST-VP7 protein by indirect enzyme-linked immunosorbent assay (ELISA) and limiting dilution. The production of the monoclonal antibody against GBRV will benefit the further study of GBRV's structures and functions and also lay a solid foundation for the research of disease prevention, clinical diagnosis, and treatment.
authors.affiliation: Hubei Key Laboratory of Genetic Regulation and Integrative Biology, College of Life Science, Huazhong Normal University , Wuhan, P.R. China .
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Deng
authors.forename: Guomei
authors.initials: G
authors.lastname: Xiong
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Cong
authors.forename: Zhonglai
authors.initials: Z
authors.lastname: Liu
authors.forename: Chao
authors.initials: C
authors.lastname: Qi
authors.forename: Jihong
authors.initials: J
authors.lastname: Yang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012401:Rotavirus / Q000145:classification / Q000276:immunology*; D012400:Rotavirus Infections / Q000175:diagnosis / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D036022:Capsid Proteins; D011993:Recombinant Fusion Proteins; C050779:VP7 protein, Rotavirus; D007544:Isopropyl Thiogalactoside; D005982:Glutathione Transferase