caa a22 4500 445312181 CHVBK 20180317142634.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s11095-011-0410-0 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0410-0 Establishing a Link Between Amino Acid Sequences and Self-Associating and Viscoelastic Behavior of Two Closely Related Monoclonal Antibodies [Elektronische Daten] [Sandeep Yadav, Alavattam Sreedhara, Sonoko Kanai, Jun Liu, Samantha Lien, Henry Lowman, Devendra Kalonia, Steven Shire] ABSTRACT: Purpose: To investigate the underlying cause for the observed differences in self-associating and viscoelastic behavior between two monoclonal antibodies, MAb1, and MAb2. Methods: Several mutants were designed by swapping charged residues in MAb1 with those present in MAb2 at their respective positions and vice versa. Rheological analysis was done at low and high shear rates. Dynamic light scattering quantified intermolecular interactions in dilute solutions; sedimentation equilibrium analysis determined the corrected weight average molecular weight (M wc) to assess the self-associating behavior in high concentration. The molecular charge was estimated from electrophoretic mobility measurements. Results: Replacing the charged residues in the CDR of MAb1 resulted in a lower M wc and solution viscosity. The corresponding changes in either just the variable light (VL) or variable heavy (VH) chain showed only a partial decrease in viscosity, whereas changes in both VL and VH chains resulted in a dramatic reduction in viscosity. The converse case where the VL and VH chains of MAb2 were made to look like MAb1 did not self-associate or show increased viscosity. Conclusions: Exposed charged residues in the CDR of MAb1 are critical in determining the self-associating and highly viscous behavior observed at high concentrations. Springer Science+Business Media, LLC, 2011 high concentration protein solution nationallicence intermolecular interactions nationallicence protein viscosity/rheology nationallicence self-association nationallicence Yadav Sandeep Department of Pharmaceutical Sciences, University of Connecticut, 06269, Storrs, Connecticut, USA aut Sreedhara Alavattam Late Stage Pharmaceutical Development, Genentech, Inc., 1 DNA Way, 94080, South San Francisco, California, USA aut Kanai Sonoko Pharmaceutical and Analytical R & D, F. Hoffmann La-Roche, Grenzacherstrasse 124, 4070, Basel, Switzerland aut Liu Jun Late Stage Pharmaceutical Development, Genentech, Inc., 1 DNA Way, 94080, South San Francisco, California, USA aut Lien Samantha Antibody Engineering, Genentech, Inc., 1 DNA way, 94080, South San Francisco, California, USA aut Lowman Henry CytomX Therapeutics, Inc., 460 Ward Drive, Suite E1, 93111, Santa Barbara, California, USA aut Kalonia Devendra Department of Pharmaceutical Sciences, University of Connecticut, 06269, Storrs, Connecticut, USA aut Shire Steven Late Stage Pharmaceutical Development, Genentech, Inc., 1 DNA Way, 94080, South San Francisco, California, USA aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/7(2011-07-01), 1750-1764 0724-8741 28:7<1750 2011 28 11095 https://doi.org/10.1007/s11095-011-0410-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0410-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yadav Sandeep Department of Pharmaceutical Sciences, University of Connecticut, 06269, Storrs, Connecticut, USA aut NATIONALLICENCE 700 1- Sreedhara Alavattam Late Stage Pharmaceutical Development, Genentech, Inc., 1 DNA Way, 94080, South San Francisco, California, USA aut NATIONALLICENCE 700 1- Kanai Sonoko Pharmaceutical and Analytical R & D, F. Hoffmann La-Roche, Grenzacherstrasse 124, 4070, Basel, Switzerland aut NATIONALLICENCE 700 1- Liu Jun Late Stage Pharmaceutical Development, Genentech, Inc., 1 DNA Way, 94080, South San Francisco, California, USA aut NATIONALLICENCE 700 1- Lien Samantha Antibody Engineering, Genentech, Inc., 1 DNA way, 94080, South San Francisco, California, USA aut NATIONALLICENCE 700 1- Lowman Henry CytomX Therapeutics, Inc., 460 Ward Drive, Suite E1, 93111, Santa Barbara, California, USA aut NATIONALLICENCE 700 1- Kalonia Devendra Department of Pharmaceutical Sciences, University of Connecticut, 06269, Storrs, Connecticut, USA aut NATIONALLICENCE 700 1- Shire Steven Late Stage Pharmaceutical Development, Genentech, Inc., 1 DNA Way, 94080, South San Francisco, California, USA aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/7(2011-07-01), 1750-1764 0724-8741 28:7<1750 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer