Improved Fluorescence Methods for High-Throughput Protein Formulation Screening. (December 2018)

• Record Type: Journal Article
• Authors: Wei, Yangjie; Larson, Nicholas R.; Angalakurthi, Siva K.; Russell Middaugh, C.
• Is Part Of: SLAS technology. Volume 23:Number 6(2018)
• Page Start: 516
• Page End: 528
• Publication Date: 2018-12
• Subjects: high-throughput -- intrinsic fluorescence -- protein formulation -- excipient screening -- melting temperature; Medical laboratory technology -- Periodicals; Laboratories -- Equipment and supplies -- Periodicals; Diagnosis, Laboratory -- Periodicals; 616.075
• DOI: 10.1177/2472630318780620
• Languages: English
• ISSNs: 2472-6303

Abstract

The goal of protein formulation development is to identify optimal conditions for long-term storage. Certain commercial conditions (e.g., high protein concentration or turbid adjuvanted samples) impart additional challenges to biophysical characterization. Formulation screening studies for such conditions are usually performed using a simplified format in which the target protein is studied at a low concentration in a clear solution. The failure of study conditions to model the actual formulation environment may cause a loss of ability to identify the optimal condition for target proteins in their final commercial formulations. In this study, we utilized a steady-state/lifetime fluorescence-based, high-throughput platform to develop a general workflow for direct formulation optimization under analytically challenging but commercially relevant conditions. A high-concentration monoclonal antibody (mAb) and an Alhydrogel-adjuvanted antigen were investigated. A large discrepancy in screening results was observed for both proteins under these two different conditions (simplified and commercially relevant). This study demonstrates the feasibility of using a steady-state/lifetime fluorescence plate reader for direct optimization of challenging formulation conditions and highlights the importance of performing formulation optimization under commercially relevant conditions.