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Cloned transchromosomic calves producing human immunoglobulin

Nature Biotechnology volume 20, pages 889–894 (2002)Cite this article

Abstract

Human polyclonal antibodies (hPABs) are useful therapeutics, but because they are available only from human donors, their supply and application is limited. To address this need, we prepared a human artificial chromosome (HAC) vector containing the entire unrearranged sequences of the human immunoglobulin (hIg) heavy-chain (H) and lambda (λ) light-chain loci. The HAC vector was introduced into bovine primary fetal fibroblasts using a microcell-mediated chromosome transfer (MMCT) approach. Primary selection was carried out, and the cells were used to produce cloned bovine fetuses. Secondary selection was done on the regenerated fetal cell lines, which were then used to produce four healthy transchromosomic (Tc) calves. The HAC was retained at a high rate (78–100% of cells) in calves and the hIg loci underwent rearrangement and expressed diversified transcripts. Human immunoglobulin proteins were detected in the blood of newborn calves. The production of Tc calves is an important step in the development of a system for producing therapeutic hPABs.

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Figure 1: Diagram of ΔHAC and ΔΔHAC construction.
Figure 2: Procedure for production of cloned Tc calves.
Figure 3: Analysis of Tc fetuses.
Figure 4: Analysis of cloned Tc calves.

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Acknowledgements

We thank J. Pommer, J. Koster, J. Molina, and D. Faber at Trans Ova Genetics for their assistance with animals and T. King and M. Nichols for their assistance in nuclear transplantation. Partial support for work in the laboratory of R.A.G. was contributed by National Science Foundation grant NSF-MCB-9986213.

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Authors and Affiliations

  1. Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., 3 Miyahara-cho, Takasaki-shi, 370-1295, Gunma, Japan

    Yoshimi Kuroiwa, Kazuma Tomizuka & Isao Ishida

  2. Hematech, LLC, 33 Riverside Avenue, Westport, 06880, CT

    Poothappillai Kasinathan, Eddie J. Sullivan, Jason G. Knott & James M. Robl

  3. Department of Biology, Amherst College, Amherst, 01002, MA

    Yoon J. Choi, Anae Duteau & Richard A. Goldsby

  4. Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, 77030, TX

    Rizwan Naeem

  5. Department of Veterinary and Animal Science University of Massachusetts, University of Massachusetts, Amherst, 01003, MA

    Barbara A. Osborne

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  1. Yoshimi Kuroiwa
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Correspondence to Isao Ishida or James M. Robl.

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Competing interests

The authors from Kirin Brewery Co. Ltd. (Y.K., K.T., and I.I) and Hematech L.L.C. (P.K., J.G.K., and J.M.R.) have personal financial interests that may be affected by publication of this article.

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Kuroiwa, Y., Kasinathan, P., Choi, Y. et al. Cloned transchromosomic calves producing human immunoglobulin. Nat Biotechnol 20, 889–894 (2002). https://doi.org/10.1038/nbt727

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