Nature Biotechnology
20, 889 - 894 (2002)
Published online: 12 August 2002; | doi:10.1038/nbt727
Cloned transchromosomic calves producing human immunoglobulinYoshimi Kuroiwa1, Poothappillai Kasinathan2, Yoon J. Choi3, Rizwan Naeem4, Kazuma Tomizuka1, Eddie J. Sullivan2, Jason G. Knott2, Anae Duteau3, Richard A. Goldsby3, Barbara A. Osborne5, Isao Ishida1
& James M. Robl21
Pharmaceutical Research Laboratory, Kirin Brewery
Co., Ltd., 3 Miyahara-cho Takasaki-shi
Gunma 370-1295, Japan. 2
Hematech, LLC, 33 Riverside
Avenue, Westport, CT 06880. 3
Department of Biology, Amherst College,
Amherst, MA 01002. 4
Department of Molecular and Human Genetics, Baylor
College of Medicine, One Baylor Plaza,
Houston, TX 77030. 5
Department of Veterinary and Animal Science,
University of Massachusetts, Amherst, MA
01003.
Correspondence should be addressed to Isao Ishida i-ishida@kirin.co.jp or James M. Robl jrobl@hematech.comHuman 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 ( )
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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