title: Monoclonal antibodies directed against chicken β2-microglobulin developed with a synthesized peptide.
abstract: We developed a panel of monoclonal antibodies (MAb) against chicken β2-microglobulin (chβ2M) by fusions between SP2/0 myeloma cells and spleen cells from mice immunized with a synthesized peptide corresponding to positions 91-119 of the COOH domain of chβ2M. Two of them, 6E7 and 3D1, identified as IgG1/κ, could react with chβ2M protein from avian macrophage HD11 cells and human 293T cells transfected with pcDNA3.1-chβ2M in immunofluorescence assays. Only a 12 kDa protein band of chβ2M could be detected in the HD11 and 293T/chβ2M cell lysates by Western blot analysis. Chicken β2M in serum and plasma could be found in Western blot by MAb 3D1. Moreover, MAb 3D1 also recognized the chβ2M antigen on the cell membranes in flow cytometry. Immunohistochemical staining with these MAbs revealed that chβ2M was present in chicken thymus, spleen, and bursa. These MAbs will be good tools for analyzing the mechanism of the chicken immune system.
authors.affiliation: Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, Yangzhou, PR China.
authors.forename: Chuan
authors.initials: C
authors.lastname: Yu
authors.forename: Qiu
authors.initials: Q
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Gao
authors.forename: Kun
authors.initials: K
authors.lastname: Qian
authors.forename: Mei
authors.initials: M
authors.lastname: Mei
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Shao
authors.forename: Gen-Hua
authors.initials: GH
authors.lastname: Wu
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Jin
authors.forename: Ai-Jian
authors.initials: AJ
authors.lastname: Qin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002060:Bursa of Fabricius / Q000378:metabolism; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens / Q000276:immunology*; D057809:HEK293 Cells; D015395:Histocompatibility Antigens Class I / Q000378:metabolism; D006801:Humans; D008264:Macrophages / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D016415:Sequence Alignment; D013154:Spleen / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism; D001613:beta 2-Microglobulin / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D015395:Histocompatibility Antigens Class I; D001613:beta 2-Microglobulin