Generation of chimeric bispecific G250/anti-CD3 monoclonal antibody, a tool to combat renal cell carcinoma

Abstract

Summary The monoclonal antibody (MAb) G250 binds to a tumour-associated antigen, expressed in renal cell carcinoma (RCC), which has been demonstrated to be a suitable target for antibody-mediated immunotherapy. A bispecific antibody having both G250 and anti-CD3 specificity can cross-link G250 antigen-expressing RCC target cells with T cells and can mediate lysis of such targets. Therapy studies with murine antibodies are limited by immune responses to the antibodies injected (HAMA response), which can be decreased by using chimeric antibodies. We generated a chimeric bispecific G250/anti CD3 MAb by transfecting chimeric genes of heavy and light chains for both the G250 MAb and the anti-CD3 MAb into a myeloma cell line. Cytotoxicity assays revealed that the chimeric bispecific MAb was capable of mediating lysis of RCC cell lines by cloned human CD8+T cells or by IL-2-stimulated peripheral blood lymphocytes (PBLs). Lysis mediated by the MAb was specific for target cells that expressed the G250 antigen and was effective at concentrations as low as 0.01 pg ml-'. The chimeric bispecific G250/anti-CD3 MAb produced may be an effective adjuvant to the currently used IL-2-based therapy of advanced renal cell arcinoma.

Hypoxia-induced carbonic anhydrase IX as a target for cancer therapy: From biology to clinical use

“…Because the highest and most frequent CA IX expression occurs in kidney cancer, this approach was systematically studied in preclinical RCC models and in clinical cohorts of RCC patients using the CA IX-specific monoclonal antibody G250 and its humanized, chimeric and bispecific variants [175][176][177][178]. This antibody was generated against a renal cell carcinoma-associated antigen G250 that was later identified as CA IX [175,179].…”

Carbonic Anhydrase IX: From Biology to Therapy

“…The immunotherapeutic approach was consistently studied in whole series of preclinical and clinical experiments in RCC models and patients with the CA IXspecific monoclonal antibody G250 and its humanized, chimeric, and bispecific variants (Oosterwijk et al 1986;Luiten et al 1996Luiten et al , 1997. This antibody was generated against an RCC-associated antigen G250 that was later identified as CA IX (Oosterwijk et al 1986;Grabmaier et al 2004).…”

Carbonic Anhydrase Inhibitors and the Management of Cancer

“…Whole series of pre-clinical and clinical experiments was performed in RCC models and patients mostly by Oostwerwijk and coworkers with the CA IX-specific monoclonal antibody G250 and its humanized, chimeric and bispecific variants [122][123][124][125][126]. CA IX has been examined also as a target for dentritic cell vaccine [127], anti-idiotype vaccine [128][129][130] and genetically modified cytotoxic cells [131,132].…”