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Production of human monoclonal antibody in eggs of chimeric chickens

Nature Biotechnology volume 23, pages 1159–1169 (2005)Cite this article

Abstract

The tubular gland of the chicken oviduct is an attractive system for protein expression as large quantities of proteins are deposited in the egg, the production of eggs is easily scalable and good manufacturing practices for therapeutics from eggs have been established. Here we examined the ability of upstream and downstream DNA sequences of ovalbumin, a protein produced exclusively in very high quantities in chicken egg white, to drive tissue-specific expression of human mAb in chicken eggs. To accommodate these large regulatory regions, we established and transfected lines of chicken embryonic stem (cES) cells and formed chimeras that express mAb from cES cell–derived tubular gland cells. Eggs from high-grade chimeras contained up to 3 mg of mAb that possesses enhanced antibody-dependent cellular cytotoxicity (ADCC), nonantigenic glycosylation, acceptable half-life, excellent antigen recognition and good rates of internalization.

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Figure 1: Tissue-restricted expression of mAb from Ov7.5mAbdns and Ov15mAbF1 vectors.
Figure 2: Deposition of mAbF1 into eggs from Ov15mAbF1 chimeras.
Figure 3: Glycosylation of mAbF1 protein produced in chicken tubular gland cells.
Figure 4: Biological activities of mAbF1 produced by chicken tubular gland cells in comparison to that produced by CHO cells.

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Acknowledgements

The work conducted at Origen Therapeutics and Texas A&M University was supported by the National Institutes of Health Small Business Innovation Research Grant GM064261. Research in the laboratory at UCLA was supported by grants AI29470, AI39187 and AI51415 from the National Institutes of Health.

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Authors and Affiliations

  1. Origen Therapeutics, 1450 Rollins Road, Burlingame, 94010, California, USA

    Lei Zhu, Marie-Cecile van de Lavoir, David F Deng, Jennifer H Diamond, Babette S Heyer, Robert M Kay, Allyn Kerchner, Philip A Leighton, Christine M Mather, Alicia Pradas-Monne, Mingxia Shi, Peggy Winters-Digiacinto, Wen Zhou & Robert J Etches

  2. Medarex Inc., 521 Cottonwood Drive, Milpitas, 95035, California, USA

    Jenny Albanese, Pina M Cardarelli, Severino Cuison, Shrikant Deshpande, Lynae Green, Edward L Halk, David B Passmore, Benjamin T Preston, Vangipuram S Rangan, Mohan Srinivasan & Susan Wong

  3. Department of Biological and Agricultural Engineering, Texas A&M University, College Station, 77843, Texas, USA

    Zivko L Nikolov & Steven G White

  4. Department of Microbiology, Immunology and Molecular Genetics and the Molecular Biology Institute, University of California Los Angeles, Los Angeles, 90095, California, USA

    David O Beenhouwer & Sherie L Morrison

  5. Division of Infectious Diseases, VA Greater Los Angeles Healthcare System, Los Angeles, 90073, California, USA

    David O Beenhouwer

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Correspondence to Robert J Etches.

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Many of the authors are employees of companies developing these technologies for commerical application.

Supplementary information

Supplementary Fig. 1

Southern blot analysis of cES clones transfected with Ov7.5 and Ov15 expression vectors. (PDF 1575 kb)

Supplementary Fig. 2

The morphology of the female chicken embryonic stem (ES) cell line 440 is shown in the upper panel. (PDF 125 kb)

Supplementary Fig. 3

cIEF data for MAbF1 produced by chicken tubular gland cells and CHO cells. (PDF 377 kb)

Supplementary Fig. 4

Thermal stabilities of chicken and CHO cell derived MAbF1. (PDF 1893 kb)

Supplementary Table 1

The extent of feather chimerism and the number of offspring obtained from male chimeras carrying cES cells derived from Barred Plymouth Rock. (PDF 31 kb)

Supplementary Table 2

Sequences for PCR primers. (PDF 37 kb)

Supplementary Table 3

Sequence analysis of chicken produced and CHO produced MAbF1 by mass spectrometry. (PDF 50 kb)

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Zhu, L., van de Lavoir, MC., Albanese, J. et al. Production of human monoclonal antibody in eggs of chimeric chickens. Nat Biotechnol 23, 1159–1169 (2005). https://doi.org/10.1038/nbt1132

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