Purification of antigenized immunoglobulins derivatized with monomethoxypolyethylene glycol

Journal of Chromatography. a
T D BrumeanuC Bona

Abstract

Genetically engineered immunoglobulins (Igs) carrying viral B or T cell peptides in the CDR3 loop, function as efficient delivery system of the defined viral epitopes. Two of these antigenized Igs (AIgs) were derivatized with 2-O-monomethoxypolyethylene glycol-4,6-dichloro-s-triazine (mPEG). Herein, we describe a two-step strategy to purify mPEG-derivatized AIgs (AIgs-mPEG). Unreacted mPEG polymers were removed by size-exclusion chromatography using ammonium hydrogencarbonate as a buffer system. Mildly PEGylated AIgs were isolated from free and highly derivatized AIgs by anion-exchange chromatography. Electrophoretic analysis indicated that the AIgs-mPEG preparation contained less than 4 x 10(-4) M unreacted mPEG. This strategy may be applied to other mPEG-derivatized monoclonal antibodies.

References

Jan 1, 1992·Biomaterials, Artificial Cells, and Immobilization Biotechnology : Official Journal of the International Society for Artificial Cells and Immobilization Biotechnology·K NhoA Abuchowski
Aug 10, 1992·Journal of Immunological Methods·C Cunningham-RundlesJ Keenan
Mar 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·N V KatreW J Laird

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Citations

Feb 5, 2011·Journal of Chromatography. a·Mark R Etzel, Tejashree Bund
Jan 23, 2010·Electrophoresis·María R PlataAngel Ríos
Feb 18, 2014·Journal of Controlled Release : Official Journal of the Controlled Release Society·David Pfister, Massimo Morbidelli
Sep 23, 2010·Pharmaceutical Development and Technology·Robert W PayneMark Cornell Manning
Mar 14, 1998·Protein Expression and Purification·Y TanR M Hoffman

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