High throughput screening for rapid and reliable prediction of monovalent antibody binding behavior in flowthrough mode

Abstract

Flowthrough (FT) anion exchange (AEX) chromatography is a widely used polishing step for the purification of monoclonal antibody (mAb) formats. To accelerate downstream process development, high throughput screening (HTS) tools have proven useful. In this study, the binding behavior of six monovalent mAbs (mvAbs) was investigated by HTS in batch binding mode on different AEX and mixed‐mode resins at process‐relevant pH and NaCl concentrations. The HTS entailed the evaluation of mvAb partition coefficients (Kp) and visualization of results in surface‐response models. Interestingly, the HTS data grouped the mvAbs into either a strong‐binding group or a weak‐binding/FT group independent of theoretical Isoelectric point. Mapping the charged and hydrophobic patches by in silico protein surface property analyses revealed that the distribution of patches play a major role in predicting FT behavior. Importantly, the conditions identified by HTS were successfully verified by 1 mL on‐column experiments. Finally, employing the optimal FT conditions (7–9 mS/cm and pH 7.0) at a mini‐pilot scale (CV = 259 mL) resulted in 99% yield and a 21–23‐fold reduction of host cell protein to <100 ppm, depending on the varying host cell protein (HCP) levels in the load. This work opens the possibility of using HTS in FT mode to accelerate downstream process development for mvAb candidates in early research.

Special Issue on “Recovery of Biological Products: Past, Present, Future”

“…The issue reflects on presentations and discussions covering state‐of‐the‐art technologies for harvesting and purifying proteins such as monoclonal antibodies (mAbs) and non‐protein modalities, such as viral vectors. Topics include: future facility designs that embrace modular (Bertran & Babi, 2024) and continuous concepts (Konoike et al, 2024; Minervini et al, 2024), novel purification technologies from fluidized bed systems and 3D‐printed monoliths (Herlevi et al, 2024; Pei et al, 2024; Saballus et al, 2024) through to new membrane and chromatography devices (Lavoie et al, 2024; Platteau et al, 2024), bioprocess development acceleration tools that embrace high throughput tools with novel sensors, data analytics, modeling, and control strategies (Andersson et al, 2024; Banerjee et al, 2024; Dürauer et al, 2024; Hahn et al, 2024; Kaufman et al, 2024; Keulen et al, 2024; Lorek et al, 2024; Sebastian et al, 2024), separation science and solutions for non‐mAb proteins (Rodriguez et al, 2024; Roush et al, 2024) and emerging viral products (Kish et al, 2024; Konstantinidis et al, 2024; Lyle et al, 2024; Perry et al, 2024; Soni et al, 2024), perspectives on the current status and future outlook for downstream processing of biological products drawn from the conference roundtable discussions (Jungbauer et al, 2024). …”

“…bioprocess development acceleration tools that embrace high throughput tools with novel sensors, data analytics, modeling, and control strategies (Andersson et al, 2024; Banerjee et al, 2024; Dürauer et al, 2024; Hahn et al, 2024; Kaufman et al, 2024; Keulen et al, 2024; Lorek et al, 2024; Sebastian et al, 2024),…”