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Polyethyleglycol Conjugation Extends the Circulatory Stability of Recombinant Human Butyrylcholinesterase
Author(s) -
Chilukuri Nageswararao,
Parikh Kalpana,
Sun Wei,
Tang Lin,
Bhupendra Doctor,
Saxena Ashima
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1141-a
Subject(s) - chemistry , monomer , peg ratio , butyrylcholinesterase , enzyme , gel permeation chromatography , recombinant dna , chromatography , affinity chromatography , biochemistry , organic chemistry , acetylcholinesterase , polymer , aché , gene , finance , economics
The therapeutic value of human serum butyrylcholinesterase (HuBChE) as a bioscavenger of organophosphate chemical warfare agents is due to its prolonged circulatory stability. Plasma HuBChE contains the tetrameric form of enzyme while the enzyme produced in vitro is a mixture of tetramers, dimers, and monomers. The goal of this study was to investigate the effect of polyethyleneglycol (PEG) conjugation on the circulatory stability of monomeric and tetrameric recombinant (r) HuBChEs. Tetrameric and monomeric rHuBChEs were produced using adenovirus expression system and purified by ammonium sulfate fractionation and affinity chromatography. Purified monomeric and tetrameric rHuBChEs were conjugated with PEG‐20K and evaluated for changes in molecular size and enzyme properties. The catalytic and inhibitory properties of monomeric and tetrameric rHuBChEs were not affected following PEG conjugation. PEG conjugation resulted in an increase in the molecular size of rHuBChEs. The mean residence time (MRT) of unmodified monomeric rHu BChE was 3 h compared to 18 h for the tetrameric rHuBChE. However, MRTs for PEG‐modified monomeric and tetrameric rHuBChEs were 31 h and 36 h respectively, which were closer to the MRT for native HuBChE (45 h). These results suggest that the circulatory stability of monomeric and tetrameric rHuBChEs can be significantly extended following PEG conjugation.