High Performance Size Exclusion Chromatography and High-Throughput Dynamic Light Scattering as Orthogonal Methods to Screen for Aggregation and Stability of Monoclonal Antibody Drug Products

Impact of process stress on protein stability in highly-loaded solid protein/PEG formulations from small-scale melt extrusion

“…Most of the analytical methods applied during protein formulation development studies focus on the investigation and characterization of proteins in solution or lyophilized proteins after their reconstitution. Analytical characterization methods for liquid protein formulations include: (i) RP-HPLC for the analysis of degradation products and chemical stability, (ii) size exclusion chromatography (SEC) for monitoring the loss of protein monomers as an indication of aggregate formation, (iii) circular dichroism (CD) and nuclear magnetic resonance (NMR) to probe conformational stability and (iv) light scattering methods for estimating the colloidal stability of protein formulations and monitoring the appearance of sub-visible particles ( Capelle et al, 2007 ; Chaudhuri et al, 2014 ; Dauer et al, 2021a ; Dauer et al, 2021b; Bhirde et al, 2020 ; Ma et al, 2020 ).…”

Evaluation and Screening of Biopharmaceuticals using Multi-Angle Dynamic Light Scattering

“…Previously, correlations between DLS and SEC have been shown by authors [11,29], though quantification of protein aggregation is hard to achieve by DLS alone. While MADLS along with orthogonal techniques has been employed for the characterization of a wide range of CQAs of adeno-associated viruses (AAVs) and lipid-based nanoparticles (LNPs) [25,30], polystyrene nanoparticles and extracellular vesicles [31], the 3-in-1 capability of MADLS for the screening of particle size, concentration and aggregation of different protein-based biopharmaceuticals has not been explored.…”

Rapid Estimation of Size-Based Heterogeneity in Monoclonal Antibodies by Machine Learning-Enhanced Dynamic Light Scattering

“…The main advantage of DLS is its ability to analyze in a wide temperature range (4–85 °C), a high-throughput approach coupled with high sensitivity whilst eliminating the need for highly trained personnel. , The significantly less consumable requirement makes DLS a significantly cheaper alternative to SEC . This explains the growing popularity of DLS studies involving protein aggregation kinetics, immunoassays, and biochemical aggregation, irrespective of the sample matrix …”

“…28,29 The significantly less consumable requirement makes DLS a significantly cheaper alternative to SEC. 30 This explains the growing popularity of DLS studies involving protein aggregation kinetics, 31 immunoassays, 32 and biochemical aggregation, 33 irrespective of the sample matrix. 34 With advancements in operating conditions, data acquisition, and optimized graphical user interface, DLS has demonstrated its potential to expand beyond limited size characterization.…”

“…45−47 Furthermore, kD has been found to be a key biophysical property for the mAbs, exhibiting statistically significant correlations with multiple biophysical attributes of antibody formulations such as apparent solubility, viscosity behavior, and electrostatic properties. 30,48 However, despite all of these advantages, DLS lacks behind SEC in the fact that PSD and percentage aggregates in the mAb are not directly correlated. 15 This is where advanced mathematical tools like artificial intelligence (AI), machine learning (ML), and neural network (NN) can be of immense help.…”