Aggregation Time Machine: A Platform for the Prediction and Optimization of Long-Term Antibody Stability Using Short-Term Kinetic Analysis

Abstract

Monoclonal antibodies are the fastest growing class of therapeutics. However, aggregation limits their shelf life and can lead to adverse immune responses. Assessment and optimization of the long-term antibody stability are therefore key challenges in the biologic drug development. Here, we present a platform based on the analysis of temperature-dependent aggregation data that can dramatically shorten the assessment of the long-term aggregation stability and thus accelerate the optimization of antibody formulations. For a set of antibodies used in the therapeutic areas from oncology to rheumatology and osteoporosis, we obtain an accurate prediction of aggregate fractions for up to three years using the data obtained on a much shorter time scale. Significantly, the strategy combining kinetic and thermodynamic analysis not only contributes to a better understanding of the molecular mechanisms of antibody aggregation but has already proven to be very effective in the development and production of biological therapeutics.

Insights into ultra-low affinity lipase-antibody noncovalent complex binding mechanisms

“…Storage of the reference lipases in their purification buffers at 4°C for 6 months resulted in the enrichment of certain glycoforms in solutions, with other glycoforms aggregating, sticking to the plastic vials, and/or precipitating. 30 Stored lipases had similar charge state distributions, implying that the tertiary structure was minimally perturbed. In all cases, refrigerated lipases failed to bind to antibodies (storage of samples at −80°C retained forms/activity) by native MS, creating a pseudo “knockout” experiment.…”

Optimization of Drug-Linker to Enable Long-term Storage of Antibody–Drug Conjugate for Subcutaneous Dosing

“…As a result, iADCs would need to have properties similar to those of the parent antibody to enable SC dosing , and also enable formulation at high concentrations of up to 100 mg/mL . During discovery, antibody screening funnels have been established to profile and select the optimal candidates with the best properties to advance , and also predict those with the characteristics likely to afford the desired long-term stability. , Such high concentrations are a challenge for monoclonal antibodies , even without the conjugation of drug-linkers that increase the hydrophobic nature of the biologic …”

“…13 During discovery, antibody screening funnels have been established to profile and select the optimal candidates with the best properties to advance 14,15 and also predict those with the characteristics likely to afford the desired long-term stability. 16,17 Such high concentrations are a challenge for monoclonal antibodies 18,19 even without the conjugation of drug-linkers that increase the hydrophobic nature of the biologic. 20 Further rationalization for the optimal design of the druglinker is supported by the solution stability data of the ADCs.…”

Discovery of ABBV-154, an anti-TNF Glucocorticoid Receptor Modulator Immunology Antibody-Drug Conjugate (iADC)

“…16 During the discovery phase antibodies are profiled in extensive screening funnels that have been established to not only enable the selection of the optimal candidates with the best properties to advance 17,18 but also forecast those with the characteristics likeliest to support prolonged liquid formulation stability, a prerequisite for subcutaneous dosing by self-administration. 19,20 High concentrations already provide a serious challenge for monoclonal antibodies 21,22 even before conjugation of drug linkers that are likely to increase the hydrophobic nature of the biologic. 23 Table 1.…”

“…This mandates that iADCs would need to have physicochemical properties similar to the parent antibody so they are compatible with subcutaneous dosing. , Even more challenging is that there would be a need to enable a high-concentration liquid formulation . During the discovery phase antibodies are profiled in extensive screening funnels that have been established to not only enable the selection of the optimal candidates with the best properties to advance , but also forecast those with the characteristics likeliest to support prolonged liquid formulation stability, a prerequisite for subcutaneous dosing by self-administration. , High concentrations already provide a serious challenge for monoclonal antibodies , even before conjugation of drug linkers that are likely to increase the hydrophobic nature of the biologic …”