Comparison of results of various methods used to determine the extent of modification of methoxy polyethylene glycol 5000-modified bovine cupri-zinc superoxide dismutase

Analytical Biochemistry
J BullockL Pachla

Abstract

The protein bovine cupri-zinc superoxide dismutase (SOD) was modified by the reaction of lysine residues with an active ester of methoxy polyethylene glycol 5000 (PEG). The extent of modification was determined by capillary zone electrophoresis, matrix-assisted laser desorption/ionization mass spectrometry, Fourier-transform infrared spectroscopy, and Raman spectroscopy and after removing PEG by alkaline hydrolysis of the linkages to SOD followed by quantification of the released PEG using gel-permeation chromatography. There was generally good agreement among the results obtained by these techniques on a typical sample of the PEG-modified protein. The results, extent of protein modification, determined by the preceding methods were compared to that found from the classical trinitrobenzene sulfonic acid (TNBS) procedure. In addition to providing alternatives to the TNBS procedure, results from the described methodologies strongly suggest that the extent of modification determined from the TNBS procedure is overestimated.

References

Jan 1, 1992·Neuropsychobiology·B ArranzJ Marcusson

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Citations

Feb 18, 2010·Journal of Drug Targeting·Guilin WangHasan Uludağ
Jun 28, 2011·Journal of Controlled Release : Official Journal of the Controlled Release Society·Elham SadeqzadehS Moein Moghimi
Nov 3, 2009·Biomaterials·Sufeng ZhangHasan Uludağ
Jul 21, 2012·Journal of Biomedical Materials Research. Part a·Stefanie M ShielsJoo L Ong
May 12, 2004·Journal of Biochemical and Biophysical Methods·Weijun Li, Zhiguo Su
Apr 26, 2007·Macromolecular Bioscience·Sufeng ZhangHasan Uludağ
Mar 17, 2007·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·Xian-Zhong LiQian Liu
Sep 23, 2010·Pharmaceutical Development and Technology·Robert W PayneMark Cornell Manning
Jun 25, 2005·Artificial Cells, Blood Substitutes, and Immobilization Biotechnology·L Arzu Bozkaya Demirel, Leman Tarhan
Jun 8, 2002·Advanced Drug Delivery Reviews·Francesco M VeroneseMauro Sergi
Jun 12, 2011·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Haining TanFengshan Wang

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