caa a22 4500 445313552 CHVBK 20180317142638.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s11095-011-0416-7 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0416-7 Aggregation Stability of a Monoclonal Antibody During Downstream Processing [Elektronische Daten] [Paolo Arosio, Giuliano Barolo, Thomas Müller-Späth, Hua Wu, Massimo Morbidelli] ABSTRACT: Purpose: To study the effect of several operative parameters, particularly pH and salt concentration, on the stability and aggregation kinetics of IgG solutions under the conditions typically encountered in downstream processing. Methods: The time evolution of the aggregates is analyzed by a combination of dynamic light scattering, size exclusion chromatography (SEC) and field flow fractionation (FFF). Secondary structure changes are monitored by circular dichroism. Results: For the given antibody, it is found that at pH lower than 4.0 addition of salt induces a reversible aggregation to oligomers accompanied by an increase in the content of the β-sheet structure. The aggregation rate increases monotonically with the salt concentration. Both the SEC and FFF techniques are successfully applied to obtain the oligomer distributions, and their results are consistent. The modified Lumry-Eyring kinetic model can well describe the time evolutions of the oligomers. Conclusions: For the given antibody, low pH and presence of salt induce conformational changes that are responsible for the reversible aggregation, but in the investigated conditions only small soluble oligomers are formed and oligomer sizes larger than trimer are negligible. For this reason, no accompanied macroscopic changes can be observed. Springer Science+Business Media, LLC, 2011 aggregation nationallicence antibody nationallicence FFF nationallicence oligomer distribution nationallicence SEC nationallicence Arosio Paolo Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut Barolo Giuliano Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut Müller-Späth Thomas Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut Wu Hua Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut Morbidelli Massimo Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/8(2011-08-01), 1884-1894 0724-8741 28:8<1884 2011 28 11095 https://doi.org/10.1007/s11095-011-0416-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0416-7 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Arosio Paolo Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut NATIONALLICENCE 700 1- Barolo Giuliano Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut NATIONALLICENCE 700 1- Müller-Späth Thomas Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut NATIONALLICENCE 700 1- Wu Hua Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut NATIONALLICENCE 700 1- Morbidelli Massimo Department of Chemistry and Applied Biosciences Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/8(2011-08-01), 1884-1894 0724-8741 28:8<1884 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer