Microscopic insight into role of protein flexibility during ion exchange chromatography by nuclear magnetic resonance and quartz crystal microbalance approaches

Journal of Chromatography. a
Dongxia HaoZhiguo Su

Abstract

Driven by the prevalent use of ion exchange chromatography (IEC) for polishing therapeutic proteins, many rules have been formulated to summarize the different dependencies between chromatographic data and various operational parameters of interest based on statically determined interactions. However, the effects of the unfolding of protein structures and conformational stability are not as well understood. This study focuses on how the flexibility of proteins perturbs retention behavior at the molecular scale using microscopic characterization approaches, including hydrogen-deuterium (H/D) exchange detected by NMR and a quartz crystal microbalance (QCM). The results showed that a series of chromatographic retention parameters depended significantly on the adiabatic compressibility and structural flexibility of the protein. That is, softer proteins with higher flexibility tended to have longer retention times and stronger affinities on SP Sepharose adsorbents. Tracing the underlying molecular mechanism using NMR and QCM indicated that an easily unfolded flexible protein with a more compact adsorption layer might contribute to the longer retention time on adsorbents. The use of NMR and QCM provided a previously unreported approa...Continue Reading

References

Sep 1, 1986·Advances in Colloid and Interface Science·W Norde
Oct 21, 1986·Biochemistry·K Gekko, Y Hasegawa
Jan 1, 1993·Biopolymers·D P Kharakoz, A P Sarvazyan
Jun 14, 2000·Journal of Chromatography. a·E HallgrenJ Ståhlberg
Jan 5, 2002·Journal of Chromatography. a·P DePhillips, A M Lenhoff
May 2, 2002·Biochimica Et Biophysica Acta·Kunihiko Gekko
Nov 26, 2003·Proceedings of the National Academy of Sciences of the United States of America·Voichita M Dadarlat, Carol Beth Post
Jan 4, 2008·The Journal of Physical Chemistry. B·Xuankuo Xu, Abraham M Lenhoff
Jan 16, 2008·Bioprocess and Biosystems Engineering·Jürgen Hubbuch, Maria Regina Kula
Aug 15, 2009·Langmuir : the ACS Journal of Surfaces and Colloids·Wai Keen ChungSteven M Cramer
Sep 15, 2010·Proceedings of the National Academy of Sciences of the United States of America·Wai Keen ChungSteven M Cramer
Mar 8, 2011·Langmuir : the ACS Journal of Surfaces and Colloids·Alexander S Freed, Steven M Cramer
Jun 22, 2011·Biophysical Journal·Alexander Steudle, Jürgen Pleiss
Aug 23, 2011·Journal of Chromatography. a·Katarzyna Wrzosek, Milan Polakovič
Jun 9, 2012·Nature Biotechnology·Uwe GottschalkAbhinav A Shukla
Jul 10, 2012·Journal of Chromatography. a·Ron GillespieSuresh Vunnum
Jan 22, 2013·Langmuir : the ACS Journal of Surfaces and Colloids·Wen-Yih ChenHsiao-Yeh-Tzu Chung Chuan
Jan 30, 2013·Biotechnology Advances·Valerie OrrC Perry Chou
Mar 19, 2014·Trends in Biotechnology·Alexander T Hanke, Marcel Ottens
Dec 30, 2014·Journal of Chromatography. a·Stewart R DodsDaniel G Bracewell
Jan 21, 2015·Journal of Chromatography. a·Linling YuYan Sun

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bacterial Cell Wall Structure (ASM)

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Bacterial Cell Wall Structure

Bacterial cell walls are made of peptidoglycan (also called murein), which is made from polysaccharide chains cross-linked by unusual peptides containing D-amino acids. Here is the latest research on bacterial cell wall structures.

Cajal Bodies & Gems

Cajal bodies or coiled bodies are dense foci of coilin protein. Gemini of Cajal bodies, or gems, are microscopically similar to Cajal bodies. It is believed that Cajal bodies play important roles in RNA processing while gems assist the Cajal bodies. Find the latest research on Cajal bodies and gems here.