Cloned transchromosomic calves producing human immunoglobulin.
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 (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.
Kuroiwa
Yoshimi
Y
Pharmaceutical Research Laboratory, Kirin Brewery Co., Ltd., 3 Miyahara-cho, Takasaki-shi, Gunma 370-1295, Japan.
Kasinathan
Poothappillai
P
Choi
Yoon J
YJ
Naeem
Rizwan
R
Tomizuka
Kazuma
K
Sullivan
Eddie J
EJ
Knott
Jason G
JG
Duteau
Anae
A
Goldsby
Richard A
RA
Osborne
Barbara A
BA
Ishida
Isao
I
Robl
James M
JM
D000818:Animals; D030801:Animals, Genetically Modified; D002417:Cattle / Q000235:genetics*; D022222:Chromosomes, Artificial, Human; D003001:Cloning, Molecular / Q000379:methods*; D015870:Gene Expression; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D005803:Genes, Immunoglobulin / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007146:Immunoglobulin lambda-Chains / Q000235:genetics; D019076:Transgenes
D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007146:Immunoglobulin lambda-Chains