title: A monoclonal antibody against human UL16-binding protein 3.
abstract: The UL16-binding proteins (ULBPs) are a novel family of human MHC class I-related, cell surface proteins that function as ligands for NKG2D. In this study, the gene encoding human ULBP3 was cloned into prokaryotic expression vector pQE30, resulting in a recombinant plasmid pQE30-ULBP3. The pQE30-ULBP3 was transformed into Escherichia coli M15 and induced the expression of recombinant protein ULBP3 (rec-ULBP3). The purified rec-ULBP3 as an antigen was used to immunize BALB/c mice. Through cell fusion, sub-cloning, and screening approach, three hybridoma cell clones expressing monoclonal antibodies (MAb) were acquired. The results from Western blot analysis, flow cytometry, and enzyme-linked immunosorbent assay showed that the hybridoma clones B2-F1-F1 and B4-C5-D11, and not G2-A4-A12, reacted with rec-ULBP3 and nature ULBP3 expressed on the cell surface of the tumor cells. In conclusion, the new MAb described here provides a valuable tool for further investigating ULBP3 function and clinical application.
authors: Department of Nuclear Medicine, The Affiliated Hospital of Jiangsu University, Zhenjiang, China. jq1001@ujs.edu.cn
authors: Chaoming
authors: C
authors: Mao
authors: Wei
authors: W
authors: Qin
authors: Deyu
authors: D
authors: Chen
authors: Guoyue
authors: G
authors: Yuan
authors: Xiao
authors: X
authors: Mou
authors: Danxia
authors: D
authors: Bao
authors: Wei
authors: W
authors: Wu
authors: Huaxi
authors: H
authors: Xu
authors: Shengjun
authors: S
authors: Wang
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis / Q000276:immunology*; D000915:Antibody Affinity; D002460:Cell Line; D015179:Colorectal Neoplasms / Q000378:metabolism; D004926:Escherichia coli; D005260:Female; D058851:GPI-Linked Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D006825:Hybridomas; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D058851:GPI-Linked Proteins; D036341:Intercellular Signaling Peptides and Proteins; D011993:Recombinant Fusion Proteins; C423229:ULBP3 protein, human