Removal of Aggregates of Monoclonal Antibodies by Ion Exchange Chromatography

The primary objective of this work was to study the
effect of resin chemistry, pH and molarity of binding and elution
buffer on aggregate removal using Cation Exchange Chromatography
and find the optimum conditions which can give efficient aggregate
removal with minimum loss of yield. Four different resins were used
for carrying out the experiments: Fractogel EMD SO3
-(S), Fractogel EMD COO-(M), Capto SP ImpRes and S Ceramic HyperD. Runs
were carried out on the AKTA Avant system. Design of Experiments
(DOE) was used for analysis using the JMP software. The
dependence of the yield obtained using different resins on the
operating conditions was studied. Success has been achieved in
obtaining yield greater than 90% using Capto SP ImpRes and
Fractogel EMD COO-(M) resins. It has also been found that a change
in the operating conditions generally has different effects on the
yields obtained using different resins.

• Ishan Arora
• Anurag S. Rathore

• Aggregates
• cation exchange chromatography
• design of experiments
• monoclonal antibodies.

[1] A. A. Shukla and J. Thomme, Trends in Biotechnology, Vol.28,
No.5.
[2] P. A. J. Rosa et al., J. Chromatogr. A 1217 (2010) 2296-2305.
[3] A. A. Shukla et al., J. Chromatogr. B 848 (2007) 28-39.
[4] Hui F. Liu, Junfen Ma, Charles Winter and Robert Bayer, Landes
Bioscience, mAbs, Vol.2 Issue5, 480-499, September/October
2010.
[5] D. Josic and Y.P. Lim: Methods for purification of antibodies,
Food technol. Biotechnol. 39(3) 215-226 (2001).
[6] J. Thommes and M. Etzel, Biotechnol. Prog. 2007, 23, 42-45.
[7] Steven M. Cramer, Venkatesh Natarajan, Ion Exchange
Chromatography, Encyclopedia of Bioprocess Technology, 2002.
[8] Feng Li, Joe X. Zhou, Xiaoming Yang, Tim Tressel and Brian Lee,
BioProcessing Journal, September/October 2005.
[9] D.C. Harris, Quantitative Chemical Analysis; 6th Edition, W.H.
Freeman and Company: New York.