High‐throughput purification tools for rapid upstream process development are interchangeable for biologics

Abstract

Upstream process development of biologics is not only productivity‐driven but also quality‐driven. Typically, most quality attributes are not directly measurable in cell culture samples due to low product concentration and purity, thus requiring some level of sample purification. As higher throughput upstream technologies become available, sample purification is becoming a bottleneck in limiting the number and types of cell culture samples that can be analyzed. The application of high‐throughput, microscale protein purification techniques has the potential to address and expand this capability. In this work, the affinity capture step of an IgG1 mAb was adapted to fit resin‐plate, resin‐tip, and mini‐column formats in an attempt to approximate the packed‐column performance by optimizing parameters such as contact time, liquid/resin ratio, and loading to produce a yield of ≥70% yield. A representative cell culture supernatant was purified using both the optimized microscale and conventional techniques, and analyzed using a comprehensive panel of product quality assays. This direct comparison demonstrated that each technique generates product of equivalent purity across a wide range of feed conditions. The analytical comparability suggests that any of the conventional and high‐throughput methods are interchangeable for biologics, allowing flexible development of an end‐to‐end integrated high‐throughput strategy.

Relationship between HETP measurements and breakthrough curves in short chromatography columns

“…dynamic binding capacity, flow nonuniformity, minicolumns, modeling, proteins 1 | INTRODUCTION Miniature columns or "minicolums" are used extensively, often in conjunction with robotic systems, to generate high-throughput data such as the dynamic binding capacity (DBC), which is critical to the design of capture steps in protein purification processes. [1][2][3][4] In order to conserve materials, such minicolumns have volumes of 0.05-1 ml with diameters of 0.5-1 cm and column lengths in the 0.25-5 cm range. 1,5,6 The small volume allows rapid process development using small amounts of protein while scouting ample ranges of load, wash, and elution conditions.…”

“…Miniature columns or “minicolums” are used extensively, often in conjunction with robotic systems, to generate high‐throughput data such as the dynamic binding capacity (DBC), which is critical to the design of capture steps in protein purification processes 1‐4 . In order to conserve materials, such minicolumns have volumes of 0.05–1 ml with diameters of 0.5–1 cm and column lengths in the 0.25–5 cm range 1,5,6 .…”

Extraction on a Robotic Platform – Autonomous Solvent Selection under Economic Evaluation Criteria

“…During the last decades, automation of the upstream section has been well established [7][8][9][10][11]. The improvements in the upstream section shifted the bottleneck towards downstream processing concerning the tremendous influence on the overall economics [3,12].…”

Flexible and Accessible Automated Operation of Miniature Chromatography Columns on a Liquid Handling Station

“…Automation is an integral part of adopting a High Throughput (HT) approach in bioprocess development activities . Robotic stations have been used to miniaturize unit operations involved frequently in the manufacture of biologics (e.g.,). Since a key part of downstream operations is product purification, and here packed bed column chromatography is the most common technique, various HT‐compatible miniaturized tools have been developed for studying and optimizing chromatographic separations .…”