Excipients Differentially Influence the Conformational Stability and Pretransition Dynamics of Two IgG1 Monoclonal Antibodies

Biophysical and formulation studies of theSchistosoma mansoniTSP-2 extracellular domain recombinant protein, a lead vaccine candidate antigen for intestinal schistosomiasis

“…These results are consistent with the results of many similar studies with other protein molecules. 25,32,33 Glycerol appeared to be a potential stabilizer; however, due to its much higher osmolarity at a comparable weight amount of sugar molecule, it was not further pursued as a stabilizer. Less effective stabilizing effects were observed for aspartic acid, possibly due to chargecharge interactions between the positive charged side chains of the antigen and the negatively charged carboxylic groups.…”

Understanding the relevance of local conformational stability and dynamics to the aggregation propensity of an IgG1 and IgG2 monoclonal antibodies

“…Detailed SLS, ANS fluorescence, CD, and DSC characterization protocols are described earlier. 16,18 Results Intrinsic Trp fluorescence (peak position and fluorescence intensity) as a function of temperature and excitation wavelength…”

“…The aggregation-prone region(s) in a threedimensional native structure of immunoglobulins can exist in regions with different degrees of solvent exposure. Such regions in immunoglobulins can manifest differences in their conformational stability [12][13][14] and dynamic [15][16][17][18][19] properties in response to changes in temperature and the presence of stabilizing or destabilizing cosolutes. 19 These variables have the potential to induce partially folded intermediate structures, which are less stable than the native state and more prone to aggregation.…”

“…Fluorescence quenching studies, used to probe protein matrix dynamics, were performed at selected temperatures including a pre-unfolding temperature range, an onset temperature (T onset ), and at the apparent thermal melting temperature (T MA ) to obtain a snapshot of local dynamics of the antibody as a function of temperature. 19 The effect of sucrose or arginine as representative stabilizing and destabilizing excipients, 18,37 respectively, were also evaluated under these conditions. 19 The current study is aimed at (1) characterizing and comparing the conformational stability and dynamic properties of regions with distinct solvent exposure at various temperatures using two additional monoclonal antibodies (an IgG1 and IgG2) and (2) understanding the relevance of local conformational stability and dynamics to the aggregation propensity of representative IgG1 and IgG2 monoclonal antibodies.…”

Control of Globular Protein Thermal Stability in Aqueous Formulations by the Positively Charged Amino Acid Excipients

“…[10][11][12][13] One such mechanism that has been proposed by multiple authors is principally based on the influence of its positively charged side chain, which is similar in structure to that of the well documented chaotrope, or protein destabiliser, guanidinium. 14,15 Similar mechanisms for both lysine and histidine do not appear to have been investigated in the literature despite the fact that molecules with structural similarity to their 50 positively charged side chains, methylamine hydrochloride and imidazole respectively, have important pharmaceutical uses themselves.…”