title: Development and characterization of new monoclonal antibodies against human recombinant CA XII.
abstract: Carbonic anhydrases (CAs) are enzymes that catalyse the reversible hydration of CO2 to bicarbonate. CA XII is considered a potential biomarker of tumor cells and a promising target for specific therapies. The aim of the current study was to develop new monoclonal antibodies (MAbs) against human recombinant CA XII and evaluate their diagnostic potential. An extracellular catalytic domain of human CA XII was expressed in E. coli and used as an immunogen. Seven stable hybridoma cell lines producing high-affinity IgG antibodies against human CA XII were generated. The majority of MAbs were highly specific to CA XII and did not cross-react with human recombinant CA I, CA II, CA VII, and CA XIII. In order to demonstrate the diagnostic value of the MAbs, they were employed for the immunohistochemistry analysis of CA XII expression in tissues. Two MAbs (15A4 and 4A6) demonstrated a strong and specific immunostaining of CA XII in human tissue specimens. Flow cytometry analysis of 5 human tumor cell lines with the MAb 15A4 revealed its immunoreactivity with cellular CA XII. In conclusion, the MAbs raised against recombinant catalytic domain of CA XII recognize cellular CA XII and represent a promising diagnostic tool for the immunodetection of CA XII-expressing cells.
authors: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 20241 Vilnius, Lithuania.
authors: Dovile
authors: 0000-0003-3547-9421
authors: D
authors: Dekaminaviciute
authors: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 20241 Vilnius, Lithuania.
authors: Rita
authors: R
authors: Lasickiene
authors: School of Medicine and Institute of Biomedical Technology, University of Tampere and Fimlab Ltd., Medisiinarinkatu 3, 33520 Tampere, Finland.
authors: Seppo
authors: S
authors: Parkkila
authors: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 20241 Vilnius, Lithuania.
authors: Vaida
authors: 0000-0001-8529-8560
authors: V
authors: Jogaite
authors: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 20241 Vilnius, Lithuania.
authors: Jurgita
authors: J
authors: Matuliene
authors: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 20241 Vilnius, Lithuania.
authors: Daumantas
authors: 0000-0002-6178-6276
authors: D
authors: Matulis
authors: Institute of Biotechnology, Vilnius University, V. A. Graiciuno 8, 20241 Vilnius, Lithuania.
authors: Aurelija
authors: 0000-0003-3122-8895
authors: A
authors: Zvirbliene
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000276:immunology*; D000912:Antibodies, Neoplasm / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D014408:Biomarkers, Tumor / Q000235:genetics / Q000276:immunology*; D002256:Carbonic Anhydrases / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D009363:Neoplasm Proteins / Q000235:genetics / Q000276:immunology*; D009369:Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000276:immunology*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D000912:Antibodies, Neoplasm; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D011994:Recombinant Proteins; D002256:Carbonic Anhydrases; C477378:carbonic anhydrase XII