title: 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.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China; Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130000, Jilin, China.
authors.forename: Lintao
authors.initials: L
authors.lastname: Song
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China.
authors.forename: Yanlin
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: School of Laboratory Medicine, Jilin Medical College, Jilin 132013, Jilin, China.
authors.forename: Huiyan
authors.initials: H
authors.lastname: Wang
authors.affiliation: Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Artur A
authors.initials: AA
authors.lastname: Belov
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China.
authors.forename: Jianlou
authors.initials: J
authors.lastname: Niu
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Shi
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China.
authors.forename: Yaoyao
authors.initials: Y
authors.lastname: Xie
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China.
authors.forename: Chaohui
authors.initials: C
authors.lastname: Ye
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China; Ministry of Education Engineering Research Center of Bioreactor and Pharmaceutical Development, Jilin Agricultural University, Changchun 130000, Jilin, China. Electronic address: xiaokunli@163.net.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Li
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China. Electronic address: nyuhuang@gmail.com.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Huang
mesh_terms: D041721:3T3-L1 Cells; D000818:Animals; D015153:Blotting, Western; D003924:Diabetes Mellitus, Type 2 / Q000188:drug therapy; D005346:Fibroblast Growth Factors / Q000737:chemistry* / Q000627:therapeutic use*; D006801:Humans; D008297:Male; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry* / Q000627:therapeutic use*
chemical_list: D011994:Recombinant Proteins; C414620:fibroblast growth factor 21; D011092:Polyethylene Glycols; D005346:Fibroblast Growth Factors
keywords: Biostability; Human fibroblast growth factor 21; Mutation; Solid-phase PEGylation; Type 2 diabetes