title: Characterization of a monoclonal antibody against CREPT, a novel protein highly expressed in tumors.
abstract: CREPT (cell-cycle related and expression-elevated protein in tumor), a novel gene also called RPRD1B and C20ORF77, was recently identified to promote tumorigenesis through up-regulation of the expression of genes related to cell cycle. The previous study demonstrated that CREPT is highly expressed in a variety of tumors and enhances the expression of Cyclin D1 by promoting the formation of a chromatin loop. To study the correlation of CREPT expression with clinical factors in different tumors, we generated a monoclonal antibody (3E10) using purified recombinant human GST-CREPT protein as an antigen. In this study, we characterized the specificity of the monoclonal antibody and cloned the gene encoding the antibody for preparation of industrial production. Our results showed that the monoclonal antibody 3E10 was sensitive and specific to recognize human endogenous CREPT protein. We have mapped the epitope of the antibody and cloned the variable region sequence of the gene encoding the antibody. We confirmed that the cloned gene produced an equivalent antibody as that produced by the original hybridoma. This study provided a basis for large-scale production of the CREPT antibody, which will be useful for the study of the role of CREPT in different tumors.
authors.affiliation: 1 State Key Laboratory of Biomembrane and Membrane Biotechnology, School of Medicine, School of Life Sciences, Tsinghua University , Beijing, China .
authors.forename: Fangli
authors.initials: F
authors.lastname: Ren
authors.forename: Ruoke
authors.initials: R
authors.lastname: Wang
authors.forename: Yanquan
authors.initials: Y
authors.lastname: Zhang
authors.forename: Chunxiao
authors.initials: C
authors.lastname: Liu
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Wang
authors.forename: Jim
authors.initials: J
authors.lastname: Hu
authors.forename: Linqi
authors.initials: L
authors.lastname: Zhang
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Chang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D018797:Cell Cycle Proteins / Q000276:immunology / Q000378:metabolism*; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000276:immunology / Q000378:metabolism*; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D018797:Cell Cycle Proteins; D009363:Neoplasm Proteins; C571225:RPRD1B protein, human; D011993:Recombinant Fusion Proteins