PMID: 11339286May 8, 2001Paper

Affinity purification of novel bispecific antibodies recognising carcinoembryonic antigen and doxorubicin

Journal of Chromatography. B, Biomedical Sciences and Applications
C H FordB G Rego

Abstract

We have developed a method which combines Protein A affinity chromatography and HPLC analytical and semipreparative hydroxyapatite affinity chromatography to purify bispecific antibodies (BsMabs) from hybrid-hybridomas secreting antibodies recognising carcinoembryonic antigen (CEA) and the chemotherapeutic drug doxorubicin (Dox). Elution of the HPLC hydroxyapatite columns with a 60-360 mM phosphate buffer gradient was found to give better separation than elution with a 60-180 mM phosphate buffer gradient. Careful monitoring of HPLC fractions by enzyme linked immunosorbent assays for anti-CEA, anti-Dox and dual anti-CEA/anti-Dox activity, and pooling of fractions on the basis of these results, enabled the purification of novel BsMabs for use in in vitro and preclinical in vivo experiments.

References

Aug 1, 2000·Immunology Today·A B van SprielJ G van De Winkel
May 15, 2001·International Journal of Cancer. Journal International Du Cancer·C H FordA Mathew

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Citations

May 15, 2001·International Journal of Cancer. Journal International Du Cancer·C H FordA Mathew
Mar 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·An WillemsNico Mertens
Jun 6, 2009·New Biotechnology·Pete Gagnon

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