A critical evaluation of self-interaction chromatography as a predictive tool for the assessment of protein–protein interactions in protein formulation development: A case study of a therapeutic monoclonal antibody

The effect of charge mutations on the stability and aggregation of a human single chain Fv fragment

“…As such a better indicator of electrostatic interactions might be a direct quantification of the protein electrostatic properties either by theoretical calculations or by experimental verification using electrophoretic measurements. The latter would be more preferred as some commonly-used buffers or salts neutralize protein electrostatic properties [40,41,46,72], which would not be reflected by a theoretical calculation. More importantly, we have provided examples where aggregation does not correlate with conformational stability and native-state protein-protein interactions.…”

“…A decreasing k D value is often correlated with increased aggregation propensity when reducing electrostatic repulsion by increasing ionic strength or using a salt or buffer ion that neutralizes protein charge [35,46,57,74,78,82,19,46,63,77,81]. Consistent with these studies, for the wild-type, SB and DSV mutants, the relative rates of aggregate growth (as reflected by the corresponding T SLS ⁄ values) and protein-protein interactions (as reflected by the k D values) decrease non-specifically upon addition of NaCl due to screening of electrostatic interactions.…”

“…The long term loss of monomer is generally predicted through quantitative monitoring of aggregation under accelerated and stress conditions over weeks to months. Recent progress has been made in: (i) the development of detailed kinetic models [2,4,5,34,42,49,60,96]; (ii) correlating aggregation kinetics with protein structure and folding [11,15,16,32,35,52,53,54,67,88]; (iii) and with native-state protein-protein interaction measurements [36,46,57,74,75,79,82].…”

The osmotic second virial coefficient for protein self-interaction: Use and misuse to describe thermodynamic nonideality

“…Despite the initial popularity of light scattering for determining second virial coefficients as a means of identifying conditions conducive to protein crystallization [8−10], the method has been largely supplanted by self-interaction chromatography [26][27][28][29][30][31][32][33][34]79] − a technique more compatible M A N U S C R I P T…”

High throughput detection of antibody self-interaction by bio-layer interferometry

“…[1][2][3][4][5] Ideally, antibodies with self-interaction tendancies could be rapidly eliminated early in the discovery stage, with minimum material consumption, to minimize downstream risks. This screening process is usually performed by low to medium throughput assays such as selfinteraction chromatography (SIC) [6][7][8][9][10][11][12][13] or cross-interaction chromatography (CIC). [14][15][16] SIC measures the retention time of an antibody as it flows across a column conjugated with the same antibody of interest (or "self").…”