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Biochemistry (Moscow)

, Volume 80, Issue 1, pp 113–119 | Cite as

PEG-chitosan and glycol-chitosan for improvement of biopharmaceutical properties of recombinant L-asparaginase from Erwinia carotovora

  • K. V. Sukhoverkov
  • E. V. KudryashovaEmail author
Article

Abstract

Conjugation with the new branched copolymers, PEG-chitosan and glycol-chitosan, is suggested to improve the therapeutic properties of L-asparaginase from Erwinia carotovora (EwA). The structure and composition of such conjugates were optimized for maximal catalytic efficiency (k cat/K M) under physiological conditions, yielding improvement by a factor of 3–6 compared to the native enzyme. This effect is attributed mainly to the shift of pH activity profile towards lower pH values due to the polycationic nature of the copolymer. The thermostability of EwA conjugates was also considerably improved. Chito-PEGylation, similarly to PEGylation, can be expected to improve pharmacokinetic properties and to reduce immunogenicity of this medically relevant enzyme. It is worth mentioning that a new versatile approach based on IR spectroscopy has been developed to determine PEG-chitosan copolymer composition as well as composition of copolymer-enzyme conjugates. The proposed analytic method is “reagent-free” and allows fast and reliable determination of parameters of interest from the single IR spectrum in contrast to laborious and unreliable methods based on polymer free amino group titration with TNBS and OPA.

Key words

recombinant L-asparaginase branched copolymers PEG-chitosan activity thermostability FTIR HPLC pH optima 

Abbreviations

DMSO

dimethyl sulfoxide

EwA

L-asparaginase from Erwinia carotovora

mPEG-suc-NHS

monomethoxypoly(ethylene glycol)-N-hydroxysuccinimidyl succinate

PEG

polyethylene glycol

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Copyright information

© Pleiades Publishing, Ltd. 2015

Authors and Affiliations

  1. 1.Chemical FacultyLomonosov Moscow State UniversityMoscowRussia

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