caa a22 4500 469073756 CHVBK 20180323132929.0 cr unu---uuuuu 170328e19920501xx s 000 0 eng 10.1007/BF01756183 doi (NATIONALLICENCE)springer-10.1007/BF01756183 Comparative biological properties of a recombinant chimeric anti-carcinoma mAb and a recombinant aglycosylated variant [Elektronische Daten] [Patricia Hand, Benjamin Calvo, Diane Milenic, Takashi Yokota, Margaret Finch, Philip Snoy, Kayhan Garmestani, Otto Gansow, Jeffrey Schlom, S. Kashmiri] Summary: It has been demonstrated previously that the degree of glycosylation of a molecule may alter its pharmacokinetic properties and, in the case of an antibody, its metabolism and other biological properties. Transfectomas producing aglycosylated chimeric B72.3(γ1) pancarcinoma monoclonal antibody (mAb) were developed by introduction of the eukaryotic expression construct pECMgpB72.3 HuG1-agly, into SP2/0 murine myeloma cells producing the chimeric κ chain of mAb B72.3. After cell cloning, one subclone with the highest binding to the TAG-72-positive human colon carcinoma was designated mAb aGcB72.3, and its biological and biochemical properties were compared with those of the chimeric B72.3(γ1), designated mAb cB72.3. Polyacrylamide gel electrophoresis showed that under non-reducing conditions, the molecular masses of the aGcB72.3 and cB72.3 mAbs were 162 kDa and 166 kDa respectively. The heavy chain of mAb aGcB72.3 had a slightly faster mobility than that of cB72.3, while the mobility of the light chains of the two chimeric mAbs was similar. No difference was observed in the isoelectric points of either chimeric mAb. Liquid competition radioimmunoassays demonstrated that the aGcB72.3 and cB72.3 mAbs have comparable binding properties to TAG-72. These studies demonstrate that aglycosylation of the chimeric IgG1 mAb B72.3 at theCh2 domain, as has been shown for other mAbs [Dorai H., Mueller B., Reisfeld R. A., Gillies S. D. (1991) Hybridoma 10:211; Morrison S. L., Oi V. T. (1989) Adv Immunol 44:65], eliminates antibody-dependent cell-mediated cytotoxicity activity, but does not substantially alter affinity or plasma clearance in mice. These studies also demonstrate for the first time (a) no difference in plasma clearance of an aglycosylated and a chimeric mAb in a primate after i.v. inoculation; (b) a difference (P ⩽0.05) in mice in the more rapid peritoneal clearance of a chimeric mAb versus an aglycosylated chimeric mAb; (c) higher (0.05 ⩽P ⩽0.1) tumor:liver ratios at 24, 72 and 168 h using111In-labeled aglycosylated chimeric mAb versus chimeric mAb. Since the liver is the major site of metastatic spread for most carcinomas, slight differences in tumor to normal liver ratios may be important in diagnostic applications. These studies thus indicate that comparative analyses of a novel recombinant construct (i.e., aglycosylated) and its standard chimeric counterpart require documentation in more than one system and are necessary if one is ultimately to define optimal recombinant/chimeric constructs for diagnosis and therapy in humans. Springer-Verlag, 1992 Chimeric anti-carcinoma mAb nationallicence Aglycosylated mAb nationallicence Biologic properties nationallicence Hand Patricia Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Calvo Benjamin Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Milenic Diane Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Yokota Takashi Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Finch Margaret Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Snoy Philip Division of Product Quality Control, Center for Biologics Evaluation and Research, Food and Drug Administration, USA aut Garmestani Kayhan Chemistry Section, Radiation Oncology Branch, National Cancer Institute, The National Institutes of Health, 20892, Bethesda, MD, USA aut Gansow Otto Chemistry Section, Radiation Oncology Branch, National Cancer Institute, The National Institutes of Health, 20892, Bethesda, MD, USA aut Schlom Jeffrey Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Kashmiri S. Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut Cancer Immunology, Immunotherapy Springer-Verlag 35/3(1992-05-01), 165-174 0340-7004 35:3<165 1992 35 262 https://doi.org/10.1007/BF01756183 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF01756183 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hand Patricia Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Calvo Benjamin Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Milenic Diane Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Yokota Takashi Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Finch Margaret Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Snoy Philip Division of Product Quality Control, Center for Biologics Evaluation and Research, Food and Drug Administration, USA aut NATIONALLICENCE 700 1- Garmestani Kayhan Chemistry Section, Radiation Oncology Branch, National Cancer Institute, The National Institutes of Health, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Gansow Otto Chemistry Section, Radiation Oncology Branch, National Cancer Institute, The National Institutes of Health, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Schlom Jeffrey Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 700 1- Kashmiri S. Laboratory of Tumor Immunology and Biology, National Cancer Institute, Building 10, Room 8B07, 9000 Rockville Pike, 20892, Bethesda, MD, USA aut NATIONALLICENCE 773 0- Cancer Immunology, Immunotherapy Springer-Verlag 35/3(1992-05-01), 165-174 0340-7004 35:3<165 1992 35 262 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer