Premium
Purification of a pyrogen‐free human growth hormone antagonist
Author(s) -
Gu Tingyue,
Zheng Yizhou,
Gu Yesong,
Haldankar Raj,
Bhalerao Nihar,
Ridgway Darin,
Wiehl Paul E.,
Chen Wen Y.,
Kopchick John J.
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260480515
Subject(s) - chemistry , antagonist , arginine , glycine , amino acid , transfection , ultrafiltration (renal) , biological activity , molecular mass , biochemistry , chromatography , gene , in vitro , receptor , enzyme
Human growth hormone (hGH) is a polypeptide with 191 amino acids and a molecular mass of 22 kilodaltons. With the aid of computer molecular simulation, an hGH analog was created by altering an hGH gene to reflect the change of one amino acid (glycine [G] 120 to arginine [R]) within the third α‐helix of the hGH molecule. This hGH analog, named hGHG120R, was found to be an hGH antagonist. It may have important implications in treating human conditions in which hGH levels are abnormally high, as found in type I diabetics. Several hundred milligrams of purified hGHG120R were needed to determine the biological activity of the antagonist in animal models. A multistep downstream process was developed to purify hGHG120R from cultured mouse L cells transfected with the hGHG120R gene. The process consisted of cell clarification, salt precipitation, membrane ultrafiltration, reversed phase high performance liquid chromatography, phase separation, and lyophiliation. This work discusses the rationale for the design of the process and experimental results on the purification of hGHG120R using the process. © 1995 John Wiley & Sons, Inc.