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Site-Specific PEGylation of Therapeutic Proteins via Optimization of Both Accessible Reactive Amino Acid Residues and PEG Derivatives

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Abstract

Modification of accessible amino acid residues with poly(ethylene glycol) [PEG] is a widely used technique for formulating therapeutic proteins. In practice, site-specific PEGylation of all selected/engineered accessible nonessential reactive residues of therapeutic proteins with common activated PEG derivatives is a promising strategy to concomitantly improve pharmacokinetics, allow retention of activity, alleviate immunogenicity, and avoid modification isomers. Specifically, through molecular engineering of a therapeutic protein, accessible essential residues reactive to an activated PEG derivative are substituted with unreactive residues provided that protein activity is retained, and a limited number of accessible nonessential reactive residues with optimized distributions are selected/introduced. Subsequently, all accessible nonessential reactive residues are completely PEGylated with the activated PEG derivative in great excess. Branched PEG derivatives containing new PEG chains with negligible metabolic toxicity are more desirable for site-specific PEGylation. Accordingly, for the successful formulation of therapeutic proteins, optimization of the number and distributions of accessible nonessential reactive residues via molecular engineering can be integrated with the design of large-sized PEG derivatives to achieve site-specific PEGylation of all selected/engineered accessible reactive residues.

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Acknowledgements

Chun Zhang and Xiao-lan Yang contributed equally to the work.

This article was supported by ‘863’ High Technology Program of China [2011AA02A108], Program for New Century Excellent Talent in University [NCET-09-0926], National Natural Science Foundation of China [no. 30672009], Natural Science Foundation Project of CQ [CSTC2011BA5039], and Chongqing Education Commission [KJ100313]. The authors have no conflicts of interest that are directly relevant to the content of this article.

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  1. Unit for Analytical Probes and Protein Biotechnology, Key Laboratory of Medical Laboratory Diagnostics of the Education Ministry, College of Laboratory Medicine, Chongqing Medical University, Daping, Yuzhong District, Chongqing, 400016, China

    Chun Zhang, Xiao-lan Yang, Yong-hua Yuan, Jun Pu &  Fei Liao

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  1. Chun Zhang
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  2. Xiao-lan Yang
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Correspondence to Fei Liao.

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Zhang, C., Yang, Xl., Yuan, Yh. et al. Site-Specific PEGylation of Therapeutic Proteins via Optimization of Both Accessible Reactive Amino Acid Residues and PEG Derivatives. BioDrugs 26, 209–215 (2012). https://doi.org/10.1007/BF03261880

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