Chitinase 3-like 2 protein monoclonal antibodies.
Chitinase 3-like 2 (CHI3L2) is one of the most overexpressed genes in glioblastoma. Despite this, both the CHI3L2 gene and its protein product CHI3L2 are poorly characterized. Here we report the generation and characterization of monoclonal antibodies to CHI3L2 protein (CHI3L2 MAbs). Bacterially expressed 6 His-tagged full-length CHI3L2 was used as antigen. Spleen cells from immunized mice were collected and fused with SP2/0 myeloma cells. Hybridoma clones 2D3 and 4D2 producing high titer CHI3L2 MAbs were identified by enzyme-linked immunosorbent assay (ELISA) and further examined for their activity with the CHI3L2 protein by Western blot analysis and immunoprecipitation. The 2D3 clone was chosen for mouse inoculation and ascites formation. Antibodies derived from the ascitic fluid specifically recognized the recombinant CHI3L2 protein and strongly interacted with CHI3L2 in glioblastoma tissue lysate, as determined by Western blot analysis. The antibodies generated may be useful as a tool in various aspects of CHI3L2 investigation.
Department of Biosynthesis of Nucleic Acids, Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 150 Zabolotnogo str.Kyiv, Ukraine.
Stanislav
S
Avdieiev
Liliia
L
Savinska
Valeriy
V
Filonenko
Vadym
V
Kavsan
D054392:Adipokines / Q000032:analysis / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D000071451:Chitinase-3-Like Protein 1; D002688:Chitinases / Q000032:analysis / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005909:Glioblastoma / Q000201:enzymology*; D006133:Growth Substances / Q000032:analysis / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D047468:Immunoprecipitation; D037102:Lectins / Q000032:analysis / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D054392:Adipokines; D000911:Antibodies, Monoclonal; C084533:CHI3L1 protein, human; D000071451:Chitinase-3-Like Protein 1; D006133:Growth Substances; D037102:Lectins; D011994:Recombinant Proteins; C571841:CHI3L2 protein, human; D002688:Chitinases