Journal of Biotechnology, 22 (1992) 283-290 283
© 1992 Elsevier Science Publishers B.V. All rights reserved 0168-1656/92/$05.00
BIOTEC 00703
A CHO strain producing high-level human IL-6
with the 3' deletion construct
Naoto Tonouchi, Naoto Koyama and Kiyoshi Miwa
Central Research Laboratories, Ajinomoto Co. Inc., Kawasaki, Japan
(Received 30 June 1991; revisionaccepted 1 August 1991)
Summary
A Chinese Hamster Ovary (CHO) strain producing high-level human inter-
leukin 6 (IL-6) was obtained, by introduction of the IL-6 cDNA from which the 3'
AU-rich region was deleted. The IL-6 mRNA of this strain was stable. The
productivity was more than 15 times higher than the previously obtained clone,
which has intact IL-6 cDNA in the same expression vector. This strain produced
IL-6 at a high level of 30/xg m1-1 in batch culture, or a continuous 20 #g per 106
ceils per day in microcarrier cultivation.
IL-6 production; CHO cells; 3' AU-rich sequence; Continuous cell culture
Introduction
AU-rich sequences are widely observed in the 3' untranslated region of many
genes including most lymphokines, interferons and protooncogenes (Shaw and
Kamen, 1986; Caput et al, 1986). It is known to be a mRNA destabilizing signal
through release of the p'oly (A) tail of the mRNA (Brewer and Ross, 1988; Wilson
and Treisman, 1988), and this sequence seems to prevent efficient lymphokine
production. IL-6, originally identified as B cell stimulatory factor-2 (BSF-2), is a
multi-potential cytokine that regulates immune response, an acute phase reaction
and hematopoiesis (Kishimoto, 1989), and cDNA cloning (Hirano et al., 1986) has
Correspondence to: N. Tonouchi, Central Research Laboratories, Ajinomoto Co. Inc., 1-1, Suzuki-cho,
Kawasaki-ku, Kawasaki 210, Japan.