Secretion of Recombinant Human Insulin-Like Growth Factor I (IGF-I)英文教材.pdfVIP

Secretion of IGF-I 149 11 Secretion of Recombinant Human Insulin-Like Growth Factor I (IGF-I) Russell A. Brierley 1. Introduction The development of efficient recombinant protein production processes can be a critical factor in whether or not a pharmaceutical therapeutic protein can enter human clinical trials and ultimately the marketplace. This is especially true for therapeutic proteins that need to be administered on a daily basis for prolonged periods or if dosage requirements are very high. The use of Pichia pastoris as a recombinant expression host strain can be an excellent choice for such situations. P. pastoris has the potential for high expression levels (1,2), efficient secretion, and proper protein folding (3–5), and is a robust fermenta- tion organism capable of high cell density on inexpensive simple basal salts medium (6). The development of an insulin-like growth factor I (IGF-I) pro- duction process in which P. pastoris is used as the recombinant host strain is discussed in this chapter. IGF-I consists of 70 amino acids with a mol wt of 7648 Dalton. This single- chain protein has three intrachain disulfide bridges. IGF-I belongs to a hetero- geneous family of peptides that share some of the biological and chemical properties of insulin. IGF-I promotes growth by mediating the effects of growth hormone. Thus, such processes as skeletal growth, cell replication, and other growth-related processes are affected by IGF-I levels. Physiological concen- trations of IGF-I have been shown to be influenced by such conditions as thy- roid disease, diabetes, and malnutrition (7). IGF-I has also been shown to act synergistically with other growth factors, such as accelerating the healing of soft and mesenchymal tissue wounds (8) and enhancing the growth of mamma- lian cells in serum-free tissue-culture medium (9). IGF-I has been indicated