Automatic Assay Preparation Platform (A2P2) for Real-Time Critical Quality Attributes Monitoring of Cell Culture...

As the biopharmaceuticals industry becomes increasingly competitive, improving speed-to-market and achieving "Right First Time" are key factors that propel a company to success. Building upon our foundational work on the Automatic Assay Preparation Platform (A2P2) initially introduced to enhance process analytical technology (PAT), this publication describes its innovative application for real-time sample preparation, acquisition, and monitoring of critical quality attributes (CQAs) for biotherapeutic production. Determination of CQA in cell culture is typically supported by analytical laboratories. Titer is determined before product purification by affinity capture column using a liquid handler. Purified product concentration is then measured before subjecting to a subsequent product quality assay. This process takes up to four separate instruments and several analysts to complete. The A2P2 system, embodying an end-to-end automated and autonomous PAT solution, is configured to streamline titer measurement, product purification, purified product concentration measurement, and CQA assay within a single run sequence using a modified ultrahigh performance liquid chromatography (UHPLC) instrument. By utilizing Chromeleon's System Suitability Tests (SST) and Intelligence Run Control (IRC) feature and the Agilent injector program, A2P2 significantly reduces analysts' hands-on time and shortens result turnaround time for improved efficiency. This evolution of the A2P2 system not only reduces laboratory footprint but also positions it as an ideal solution for real-time bioprocess monitoring of CQA, further advancing our commitment to accelerating the delivery of safe and effective biotherapeutics.