Exploitation of a monoclonal antibody for weak affinity-based separation in capillary gel electrophoresis

Electrophoresis
H LjungbergS Nilsson

Abstract

Weak biospecific recognition has been established for affinity separation in high performance liquid chromatography (HPLC). The use of weak affinity chromatography (WAC) has been limited previously by the insufficient separation efficiency achieved, allowing only some 1000 plates/m to be obtained. However, it has been shown that chiral drug separation can be performed with capillary affinity gel electrophoresis (CAGE) at considerably improved efficiency as compared with traditional chromatographic procedures. Our present study demonstrates the potential of weak affinity monoclonal antibodies as a generic method for immunologically based separations in capillary electrophoresis. Monoclonal antibodies were polymerized within a silica capillary and were used for the separation of structurally similar carbohydrate antigens. The results indicate that weak biospecific interactions can be utilized in a CAGE format to produce highly selective separation of the alpha- and beta-forms of p-nitrophenyl-labeled maltose. It remains to be seen, however, how efficient weak affinity separation in CAGE can be compared with affinity HPLC protocols. Details of typical separations and of the preparation of the antibody gel are presented.

References

Feb 15, 1988·Analytical Biochemistry·S OhlsonD Zopf
Mar 30, 1984·Journal of Immunological Methods·A LundbladD Zopf
Jan 17, 1997·Journal of Chromatography. a·S OhlsonA Lundblad

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Citations

Mar 17, 1999·Journal of Molecular Recognition : JMR·L LeicktS Ohlson
Aug 6, 1998·Journal of Immunological Methods·M StrandhS Ohlson
Dec 5, 1998·Journal of Immunological Methods·L LeicktS Ohlson
Feb 1, 2000·Trends in Biotechnology·S OhlsonC F Mandenius
Apr 10, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Niels H H Heegaard, Robert T Kennedy
Oct 29, 1998·Journal of Chromatography. B, Biomedical Sciences and Applications·N H HeegaardN A Guzman
Jun 12, 2009·Electrophoresis·Niels H H Heegaard
Nov 30, 2011·Analytica Chimica Acta·Zaifang ZhuShaorong Liu
Apr 4, 2006·Journal of Immunological Methods·Reine JohanssonSten Ohlson
Aug 29, 2006·Analytical Biochemistry·Henrik A EngströmSten Ohlson
Jul 6, 2000·Analytical Chemistry·S N Krylov, N J Dovichi

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