caa a22 4500 469115122 CHVBK 20180323133122.0 cr unu---uuuuu 170328e19920601xx s 000 0 eng 10.1007/BF00780762 doi (NATIONALLICENCE)springer-10.1007/BF00780762 Application of chemically desialylated and degalactosylated human glycophorin for induction and characterization of anti-Tn monoclonal antibodies [Elektronische Daten] [Maria Duk, Iga Steuden, Danuta Dus, Czeslaw Radzikowski, Elwira Lisowska] Human erythrocyte glycophorin was desialylated by mild acid hydrolysis and degalactosylated by Smith degradation. Two monoclonal antibodies (Tn5 and Tn56) obtained by immunization of mice with this ‘artificial' Tn antigen were characterized and compared in some experiments with two antibodies (BRIC111 and LM225) obtained in other laboratories by immunization with Tn erythrocytes. The specific binding of the antibodies to glycophorins desialylated and degalactosylated on the nitrocellulose blot and to asialo-agalactoglycophorin-coated ELISA plates, and reactions with authentic Tn antigen served for identification of their anti-Tn specificity. The antibodies were further characterized in inhibition assay with various glycoproteins. The antibody Tn5 (similar to BRIC111) was shown to be specific for human erythrocyte Tn antigen, whereas Tn56 reacted strongly with different glycoproteins carrying O-linked GalNAcα- residues, and was strongly bound to the murine adenocarcinoma cell line Ta3-Ha. The antibodies Tn5, Tn56 and BRIC111 were similarly inhibited by ovine submaxillary mucin (OSM) and asialoOSM, but the antibody LM225 showed a distinct preference in reaction with OSM (sialosyl-Tn antigen). The results show that Tn antigen, obtained by chemical modifications of human glycophorin, enables the preparation and characterization of anti-Tn monoclonal antibodies, without using rare Tn erythrocytes. Chapman & Hall, 1992 Glycophorin nationallicence Smith degradation nationallicence Tn nationallicence sialosyl-Tn nationallicence monoclonal antibodies nationallicence HuGph : human erythrocyte glycophorin nationallicence HoGph : horse erythrocyte glycophorin nationallicence OSM : ovine submaxillary mucin nationallicence mAb : monoclonal antibody nationallicence ELISA : enzyme-linked immunosorbent assay nationallicence SDS-PAGE : sodium dodecylsulfate polyacrylamide gel electrophoresis nationallicence PBS : phosphate buffered saline (0.01m Na2HPO4/0.15m NaCl, pH 7.2) nationallicence BSA : bovine serum albumin nationallicence TBS : 0.05m Tris-HCl/0.15m NaCl, pH 7.2 nationallicence TGr : transformation grade nationallicence Duk Maria Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut Steuden Iga Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut Dus Danuta Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut Radzikowski Czeslaw Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut Lisowska Elwira Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut Glycoconjugate Journal Kluwer Academic Publishers 9/3(1992-06-01), 148-153 0282-0080 9:3<148 1992 9 10719 https://doi.org/10.1007/BF00780762 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00780762 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Duk Maria Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut NATIONALLICENCE 700 1- Steuden Iga Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut NATIONALLICENCE 700 1- Dus Danuta Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut NATIONALLICENCE 700 1- Radzikowski Czeslaw Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut NATIONALLICENCE 700 1- Lisowska Elwira Department of Tumour Immunology, Institute of Immunology and Experimental Therapy, 53-114, Wrocław, Poland aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 9/3(1992-06-01), 148-153 0282-0080 9:3<148 1992 9 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer