A solid-phase PEGylation strategy for protein therapeutics using a potent FGF21 analog.

Abstract

Fibroblast growth factor 21 (FGF21) is an endocrine-acting hormone that has the potential to treat metabolic diseases, such as type 2 diabetes and obesity. Development of FGF21 into a therapeutic has been hindered due to its low intrinsic bio-stability, propensity towards aggregation and its susceptibility to in vivo proteolytic degradation. In order to address these shortcomings, we've developed recombinant human FGF21 variants by strategically introducing cysteine residues via site-directed mutagenesis, and have also developed a solid-phase nickel affinity PEGylation strategy, whereby engineered, surface-exposed cysteine residues of immobilized proteins were used as a platform to efficiently and site-selectively conjugate with PEG-maleimide. The engineered PEGylated FGF21 conjugates retained its biological functions, as well as demonstrated an increase in half-life by over 211.3 min. By demonstrating the biological activity of the FGF21 analog as a prototype, we have also provided a "generalized" solid-phase approach to effectively increase serum half-life of protein therapeutics.

• Authors: Lintao Song, Yanlin Zhu, Huiyan Wang, Artur A Belov, Jianlou Niu, Lu Shi, Yaoyao Xie, Chaohui Ye, Xiaokun Li, Zhifeng Huang
• Journal: Biomaterials (Biomaterials) Vol. 35 Issue 19 Pg. 5206-15 (Jun 2014) ISSN: 1878-5905 [Electronic] Netherlands
• PMID: 24685265 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2014 Elsevier Ltd. All rights reserved.

Chemical References

• Recombinant Proteins
• fibroblast growth factor 21
• Polyethylene Glycols
• Fibroblast Growth Factors

Topics

• 3T3-L1 Cells
• Animals
• Blotting, Western
• Diabetes Mellitus, Type 2 (drug therapy)
• Fibroblast Growth Factors (chemistry, therapeutic use)
• Humans
• Male
• Mice
• Polyethylene Glycols (chemistry)
• Rats
• Recombinant Proteins (chemistry, therapeutic use)