Development of ABX-EGF, a fully human anti-EGF receptor monoclonal antibody, for cancer therapy

doi:10.1016/S1040-8428(00)00134-7

Critical Reviews in Oncology/Hematology

Volume 38, Issue 1, April 2001, Pages 17-23

https://doi.org/10.1016/S1040-8428(00)00134-7 Get rights and content

Abstract

Overexpression of epidermal growth factor receptor (EGFr) has been demonstrated on many human tumors, and the increase in receptor expression levels has been linked with a poor clinical prognosis. Blocking the interaction of EGFr and the growth factors could lead to the arrest of tumor growth and possibly result in tumor cell death. To this end, using XenoMouse™ technology, ABX-EGF, a human IgG2 monoclonal antibody (mAb) specific to human EGFr, has been generated. ABX-EGF binds EGFr with high affinity (5×10⁻¹¹ M), blocks the binding of both EGF and transforming growth factor-α (TGF-α) to various EGFr-expressing human carcinoma cell lines, and inhibits EGF-dependent tumor cell activation, including EGFr tyrosine phosphorylation, increased extracellular acidification rate, and cell proliferation. In vivo ABX-EGF prevents completely the formation of human epidermoid carcinoma A431 xenografts in athymic mice. More importantly, administration of ABX-EGF without concomitant chemotherapy results in complete eradication of established tumors. No tumor recurrence was observed for more than 8 months following the last antibody injection, further indicating complete tumor cell elimination by the antibody. Inhibition of human pancreatic, renal, breast and prostate tumor xenografts which express different levels of EGFr by ABX-EGF was also achieved. Tumor expressing more than 17 000 EGFr molecules per cell showed significant growth inhibition when treated with ABX-EGF. ABX-EGF had no effect on EGFr-negative tumors. The potency of ABX-EGF in eradicating well-established tumors without concomitant chemotherapy indicates its potential as a monotherapeutic agent for treatment of multiple EGFr-expressing human solid tumors, including those where no effective chemotherapy is available. Utilization of mAbs directed to growth factor receptors as cancer therapeutics has been validated recently by the tumor responses obtained from clinical trials with Herceptin, the humanized anti-HER2 antibody, in patients with HER2 overexpressing metastatic breast cancer. Being a fully human antibody, ABX-EGF is anticipated to exhibit a long serum half-life and minimal immunogenicity with repeated administration, even in immunocompetent patients. These results demonstrate the potent anti-tumor activity of ABX-EGF and its therapeutic potential for the treatment of multiple human solid tumors that overexpress EGFr.

Section snippets

Evolution of monoclonal antibody technology

With the development of hybridoma technology by Kohler and Milstein more than 25 years ago, the potential of monoclonal antibodies (mAbs) for human therapy was easily appreciated [1]. However, the promise of the ‘magic bullet’ was not immediately realized since the first generation of mAbs was derived from mouse and the mouse mAbs were in general immunogenic in humans leading to the generation of human anti-mouse antibody (HAMA) responses, thus limiting efficacy in long term and repeated

EGFr overexpression on human tumors

Various growth factors and growth factor receptors have been implicated in a wide variety of human cancers. EGF is a mitogenic hormone that regulates the proliferation and differentiation of normal and neoplastic cells in vitro as well as in vivo. Production of TGF-α, an EGF-related polypeptide growth factor, has been associated with cellular transformation. The EGFr, which binds both EGF and TGF-α, is structurally related to the transforming protein encoded by the v-erbB oncogene of avian

Conclusion

XenoMouse technology has been proven to be a useful tool to develop human therapeutic mAbs for the treatment of variety of diseases, including cancer and inflammatory diseases [21], [24]. The XenoMouse-derived fully human anti-EGFr mAb, ABX-EGF, demonstrated potent anti-tumor activity as a monotherapeutic agent in a variety of human carcinoma xenografts that overexpress EGFr. The observation that tumor growth inhibition by ABX-EGF is dependent on EGFr number suggests that the growth of EGFr

Reviewers

This article was reviewed by Dr Robert B. Cohen, Division of Hematology/Oncology, Box 800716, University of Virginia Health System, Charlottesville, VA 22908, USA; Shimon Slavin, MD, Cancer Immunotherapy Research Center, Hadassah University Hospital, Jerusalem 91120, Israel; Gerd Ritter, Ph.D., Associate Member, Lundwig Institute for Cancer Research, New York Branch @ MSKCC, 1275 York Avenue, New York, NY 10021, USA.

Xiao-Dong Yang is Senior Scientist and the head of the Preclinical Biology Division at Abgenix, Inc.

References (24)

D.S. Salomon et al.
Epidermal growth factor-related peptides and their receptors in human malignancies
Crit. Rev. Oncol. Hematol.
(1995)
P.P. Di Fiore et al.
Overexpression of the human EGF receptor confers an EGF-dependent transformed phenotype to NIH 3T3 cells
Cell
(1987)
D.E. Neal et al.
Epidermal-growth-factor receptors in human bladder cancer: comparison of invasive and superficial tumours
Lancet
(1985)
J. Baselga et al.
Receptor blockade with monoclonal antibodies as anti-cancer therapy
Pharmacol. Ther.
(1994)
G. Kohler et al.
Continuous cultures of fused cells secreting antibody of predefined specificity
Nature
(1975)
S. Morrison et al.
L. Riechmann et al.
Reshaping human antibodies for therapy
Nature
(1988)
A. Jakobovits et al.
Analysis of homozygous mutant chimeric mice: deletion of the immunoglobulin heavy-chain joining region blocks B-cell development and antibody production
Proc. Natl. Acad. Sci. USA
(1993)
L.L. Green et al.
Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs
Nat. Genet.
(1994)
A. Jakobovits et al.
Germ-line transmission and expression of a human-derived yeast artificial chromosome
Nature
(1993)

M.J. Mendez et al.

Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response

Nat. Genet.

(1997)

L.L. Green et al.

Regulation of B cell development by variable gene complexity in mice reconstituted with human immunoglobulin yeast artificial chromosomes

J. Exp. Med.

(1998)

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Xiao-Dong Yang is Senior Scientist and the head of the Preclinical Biology Division at Abgenix, Inc.

View full text

Development of ABX-EGF, a fully human anti-EGF receptor monoclonal antibody, for cancer therapy

Abstract

Section snippets

Evolution of monoclonal antibody technology

EGFr overexpression on human tumors

Conclusion

Reviewers

Crit. Rev. Oncol. Hematol.

Cell

Lancet

Pharmacol. Ther.

Continuous cultures of fused cells secreting antibody of predefined specificity

Nature

Reshaping human antibodies for therapy

Nature

Analysis of homozygous mutant chimeric mice: deletion of the immunoglobulin heavy-chain joining region blocks B-cell development and antibody production

Proc. Natl. Acad. Sci. USA

Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs

Nat. Genet.

Germ-line transmission and expression of a human-derived yeast artificial chromosome

Nature

Functional transplant of megabase human immunoglobulin loci recapitulates human antibody response

Nat. Genet.

Regulation of B cell development by variable gene complexity in mice reconstituted with human immunoglobulin yeast artificial chromosomes

J. Exp. Med.