title: Production of rabbit monoclonal antibodies against mouse embryonic stem cells and identification of pluripotency-associated surface antigens.
abstract: Embryonic stem (ES) cells are pluripotent stem cells derived from the inner cell mass of the blastocyst. ES cell surface molecules are important for the identification, labeling, sorting, quality control and functional studies of ES cells. Currently, knowledge of ES surface molecules is limited. To identify new surface molecules, we generated a panel of rabbit monoclonal antibodies (rMabs) against mouse ES (mES) cells. We identified three monoclonal antibodies that interact with molecules on the mES cell surface and found that the expression of their respective antigens decreased upon mES cell differentiation. The antigen of the rMab ZjuESrMab29 was identified as granulocyte macrophage colony-stimulating factor receptor α (GM-CSFR α) by liquid chromatography-tandem mass spectrometry (LC-MS/MS). This study demonstrated that rabbit monoclonal antibody production via whole-cell immunization could be a practical method for the discovery of stem cell surface antigens.
authors.affiliation: Institute of Cell Biology and Genetics, Zhejiang University, Hangzhou, Zhejiang Province, China.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Tan
authors.forename: Jiarong
authors.initials: J
authors.lastname: Zhang
authors.forename: Zhongyuan
authors.initials: Z
authors.lastname: Su
authors.forename: Bin
authors.initials: B
authors.lastname: Gu
authors.forename: Xinhang
authors.initials: X
authors.lastname: Jiang
authors.forename: Jingfeng
authors.initials: J
authors.lastname: Luo
authors.forename: Huijiao
authors.initials: H
authors.lastname: Ji
authors.forename: Guangchuan
authors.initials: G
authors.lastname: Wang
authors.forename: Bo
authors.initials: B
authors.lastname: Tao
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Zhao
authors.forename: Liangbiao
authors.initials: L
authors.lastname: Chen
authors.forename: Guoliang
authors.initials: G
authors.lastname: Yu
authors.forename: Weimin
authors.initials: W
authors.lastname: Zhu
authors.forename: Ming
authors.initials: M
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D000954:Antigens, Surface / Q000276:immunology* / Q000302:isolation & purification; D002454:Cell Differentiation / Q000276:immunology; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology / Q000276:immunology*; D000939:Epitopes / Q000276:immunology / Q000302:isolation & purification; D005434:Flow Cytometry; D006825:Hybridomas / Q000276:immunology; D007114:Immunization / Q000379:methods; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000276:immunology*; D011817:Rabbits; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor / Q000276:immunology; D053719:Tandem Mass Spectrometry
chemical_list: D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000939:Epitopes; D016187:Receptors, Granulocyte-Macrophage Colony-Stimulating Factor