PMID: 9433462Jan 20, 1998Paper

Single-step purification of bispecific monoclonal antibodies for immunotherapeutic use by hydrophobic interaction chromatography

Journal of Immunological Methods
O ManzkeH Bohlen

Abstract

A method for large scale production and single-step purification of bispecific antibodies is described. Hybrid-hybridomas were grown in hollow-fibre bioreactors with an average yield of 8 to 12 g of immunoglobulin per month. Bispecific antibodies were purified from the bioreactor supernatant by hydrophobic interaction chromatography which resolves bispecific antibodies, monospecific immunoglobulins, and culture medium supplements in one single chromatographic step. Proteins were analyzed by ELISA, SDS-PAGE, isoelectric focussing, indirect fluorescence staining, CTL-stimulation and T-cell proliferation assays. Finally, antibody preparations were checked for the presence of endotoxin and mouse DNA. Our results suggest that functional bispecific antibodies for use in therapeutic applications can be batch purified from bioreactor harvest by hydrophobic interaction chromatography in a single step. Compared to other methods such as affinity chromatography (protein A/G), ion-exchange or hydroxyapatite chromatography, our protocol offers a substantial reduction in labor time, cost, protein loss, and risk of contamination.

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Citations

Mar 17, 2001·International Journal of Cancer. Journal International Du Cancer·O ManzkeH Bohlen
Feb 28, 2001·Journal of Immunological Methods·D R Husereau, M R Suresh
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·E Boschetti
Jun 29, 2002·Journal of Biochemical and Biophysical Methods·Seema Gupta, Mavanur Suresh
Jul 20, 2002·Trends in Biotechnology·Egisto Boschetti
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·An WillemsNico Mertens
Sep 11, 2004·Protein Expression and Purification·Mark C MowryAnu Subramanian
Nov 18, 2003·Clinical and Experimental Immunology·M JensenF Berthold
Jan 9, 2019·MAbs·Gunasekaran DhandapaniGuy Patchornik
Dec 7, 2020·New Biotechnology·Gunasekaran DhandapaniGuy Patchornik
Jun 1, 2021·Antibody Therapeutics·Serene W Chen, Wei Zhang
Dec 6, 2016·Advanced Drug Delivery Reviews·Zehra ElgundiVeysel Kayser

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