PMID: 7537104Apr 13, 1995Paper

Receptor-ligand interactions measured by an improved spun column chromatography technique. A high efficiency and high throughput size separation method

Biochimica Et Biophysica Acta
Soren BuusL O Pedersen

Abstract

Size exclusion chromatography may under the right circumstances be an easy and powerful way to measure in solution the interaction between a receptor an dits ligand. Spun column chromatography is a fast size exclusion technique of increasing popularity, however, little information exists on the method development essential to obtain efficient separation in particular when used for analytical purposes. In this paper we describe a systematic approach to select the optimal parameters for spun column separation including a simple modification of the technique whereby the spun columns are eluted by high-speed gradient centrifugation. This modification is easy to implement and it considerably improves spun column performance. We hypothesize that the high-speed centrifugation step leads to the release of additional buffer which assists in the complete elution of excluded molecules while the gradient centrifugation helps to achieve equilibrium across the gel matrix during the elution. The new method has been used successfully for several different receptor-ligand interactions, and this paper describes a general approach on how to develop new applications of the technique.

References

Aug 1, 1977·Analytical Biochemistry·M Krieger, J Tolber
Sep 1, 1973·Analytical Biochemistry·M W Neal, J R Florini

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Citations

Apr 28, 2007·Cancer Immunology, Immunotherapy : CII·Mette ThornMogens H Claesson
Jul 4, 1998·Scandinavian Journal of Immunology·A StraumforsS Buus
Jan 25, 2003·Scandinavian Journal of Immunology·M L M AndersenM H Claesson
Sep 17, 1996·Proceedings of the National Academy of Sciences of the United States of America·A StryhnJ Engberg
Dec 5, 2006·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Thomas BlicherMichael Gajhede
Jun 10, 2008·Acta Crystallographica. Section F, Structural Biology and Crystallization Communications·Gustav RøderMichael Gajhede
Nov 12, 2010·Immunome Research·Claus LundegaardMorten Nielsen
Apr 24, 2009·Protein Science : a Publication of the Protein Society·Sune F L JustesenSøren Buus
Aug 21, 2010·Immunology·Namrata Tomar, Rajat K De
Apr 30, 2003·Protein Science : a Publication of the Protein Society·Morten NielsenOle Lund
Jul 28, 2005·Protein Science : a Publication of the Protein Society·Henrik FerréSøren Buus
Mar 29, 2002·The Journal of Investigative Dermatology·Lars Østergaard PedersenJürgen C Becker
Jan 30, 2004·The Journal of Biological Chemistry·Burkhard FleckensteinLudvig M Sollid
Feb 9, 2013·Current Protocols in Immunology·John SidneyAlessandro Sette
Apr 25, 2008·Current Protocols in Immunology·S BuusL Østergaard Pedersen
Apr 4, 2001·Scandinavian Journal of Immunology·T R PetersenM H Claesson
Jul 22, 2004·The Journal of Immunology : Official Journal of the American Association of Immunologists·Shuo-Wang QiaoLudvig M Sollid

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