title: Development and characterization of domain-specific monoclonal antibodies produced against human SLAMF9.
abstract: SLAMF9 is a member of the signaling lymphocyte-activating molecule (SLAM) immunoreceptor family. The SLAM family receptors are expressed in a broad range of immune cells and play an important role in immunity. To date, SLAMF9 is the least studied member of this family. Its ligand, signaling properties, and cells on whose surface it is expressed are unknown. We generated hybridoma clones 6E11 and 7G5 secreting monoclonal antibodies specific to human SLAMF9. BALB/c mice were immunized with Escherichia coli-expressed purified SLAMF9 protein; splenocytes from these mice were fused with mouse myeloma cell line NS-1. Based on isotyping of the MAbs, clone 6E11 was referred to the IgG1 subclass, while 7G5 to IgG2b. The specificity of these MAbs was assessed by ELISA, immunoblotting, immunohistochemistry, and flow cytometry. According to the results of epitope analysis, clone 6E11 reacts with the C2-like domain, whereas 7G5 is specific to the V-like domain of the SLAMF9 molecule. The generated MAbs were demonstrated to be applicable in various immunochemical analyses. They may be useful tools in studies clarifying the expression and function of human SLAMF9.
authors.affiliation: 1 Institute of Molecular and Cellular Biology , Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia .
authors.forename: Olga
authors.initials: O
authors.lastname: Volkova
authors.forename: Sergey
authors.initials: S
authors.lastname: Guselnikov
authors.forename: Ludmila
authors.initials: L
authors.lastname: Mechetina
authors.forename: Nicolai
authors.initials: N
authors.lastname: Chikaev
authors.forename: Konstantin
authors.initials: K
authors.lastname: Baranov
authors.forename: Sergey
authors.initials: S
authors.lastname: Kulemzin
authors.forename: Evdokiya
authors.initials: E
authors.lastname: Reshetnikova
authors.forename: Alexander
authors.initials: A
authors.lastname: Najakshin
authors.forename: Alexander
authors.initials: A
authors.lastname: Taranin
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology*; D000918:Antibody Specificity; D015703:Antigens, CD / Q000235:genetics* / Q000276:immunology*; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D005434:Flow Cytometry; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D017434:Protein Structure, Tertiary / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000276:immunology*; D000071176:Signaling Lymphocytic Activation Molecule Family; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1
chemical_list: D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D011956:Receptors, Cell Surface; C000604061:SLAMF9 protein, human; D000071176:Signaling Lymphocytic Activation Molecule Family; C000604064:Slamf9 protein, mouse; D000071177:Signaling Lymphocytic Activation Molecule Family Member 1