Purification of antigenized immunoglobulins derivatized with monomethoxypolyethylene glycol.
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.
Brumeanu
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TD
Department of Microbiology, Mount Sinai School of Medicine, New York, NY 10029, USA.
Zaghouani
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H
Bona
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D005450:Fluorescamine / Q000032:analysis; D005818:Genetic Engineering; D018796:Immunoconjugates / Q000276:immunology / Q000302:isolation & purification*; D007136:Immunoglobulins / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D011092:Polyethylene Glycols / Q000737:chemistry*; D051381:Rats
D000911:Antibodies, Monoclonal; D000939:Epitopes; D018796:Immunoconjugates; D007136:Immunoglobulins; D005450:Fluorescamine; D011092:Polyethylene Glycols; C028210:monomethoxypolyethylene glycol