Carboxyalkylated Histidine is a PH-dependent Product of Pegylation with SC-PEG

Overview

Journal Pharm Res

Specialties Pharmacology
Pharmacy

Date 2001 Oct 31

PMID 11683252

Citations 3

Authors

D C WYLIE

M Voloch

S Lee

Y H Liu

C Cutler

B Pramanik

Affiliations

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Abstract

Purpose: Pegylation of therapeutic protein usually results in a mixture of monopegylated proteins with differing sites of modification. With rh-interferon-alpha2A pegylation, we have found that this heterogeneity includes two classes of pegylation site chemistry, the relative proportions of which can be adjusted by reaction pH.

Methods: The effect of pegylation reaction pH on the relative proportion of three peaks produced was investigated. Products were purified and characterized by peptide mapping, chemical stability to neutral hydroxylamine, and biologic activity.

Results: Reactions at basic pH levels produced a mixture of products pegylated at lysine residues as has been observed elsewhere. However, the dominant product of reactions at mildly acidic levels of pH showed distinct chemistry and higher cytopathic effect activity. The primary site of modification at this pH was His34. We developed a quantitative assay using sensitivity to neutral hydroxylamine to measure the proportion of urethane bonds involving carboxyalkylated histidines. This assay showed that histidine was pegylated preferentially at low pH levels with another protein, rh-Interleukin-10.

Conclusions: Reaction pH can be used to select the preferred pegylation site chemistry.

Citing Articles

Research progress on the PEGylation of therapeutic proteins and peptides (TPPs).

Li C, Li T, Tian X, An W, Wang Z, Han B Front Pharmacol. 2024; 15:1353626.

PMID: 38523641 PMC: 10960368. DOI: 10.3389/fphar.2024.1353626.

Lysine-PEGylated Cytochrome C with Enhanced Shelf-Life Stability.

Santos J, Feitosa V, Meneguetti G, Carretero G, Coutinho J, Ventura S Biosensors (Basel). 2022; 12(2).

PMID: 35200354 PMC: 8869816. DOI: 10.3390/bios12020094.

Effects of localized interactions and surface properties on stability of protein-based therapeutics.

Mills B, Laurence Chadwick J J Pharm Pharmacol. 2016; 70(5):609-624.

PMID: 27861887 PMC: 5425321. DOI: 10.1111/jphp.12658.

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