Applying high‐throughput methods to develop a purification process for a highly glycosylated protein

Abstract

Micro-scale chromatography formats are becoming more routinely used in purification process development because of their ability to rapidly screen large number of process conditions at a time with minimal material. Given the usual constraints that exist on development timelines and resources, these systems can provide a means to maximize process knowledge and process robustness compared to traditional packed column formats. In this work, a high-throughput, 96-well filter plate format was used in the development of the cation exchange and hydrophobic interaction chromatography steps of a purification process designed to alter the glycoform distribution of a small protein. The significant input parameters affecting process performance were rapidly identified for both steps and preliminary operating conditions were identified. These ranges were verified in a packed chromatography column in order to assess the ability of the 96-well plate to predict packed column performance. In both steps, the 96-well plate format consistently led to underestimated glycoform-enrichment levels and to overestimated product recovery rates compared to the column-based approach. These studies demonstrate that the plate format can be used as a screening tool to narrow the operating ranges prior to further optimization on packed chromatography columns.

Separation of recombinant antibodies from DNA using divalent cations

“…High-throughput methods enable the rapid screening of suitable operation parameters [23][24][25][26]; although these methods are more popular for establishing chromatography methods, for nonchromatography methods it is simple to use high-throughput microtiter-plate technology to find suitable conditions. High-throughput methods enable the rapid screening of suitable operation parameters [23][24][25][26]; although these methods are more popular for establishing chromatography methods, for nonchromatography methods it is simple to use high-throughput microtiter-plate technology to find suitable conditions.…”

Lessons learned in building high‐throughput process development capabilities

“…Descriptions of scale down chromatography studies have been well described in the literature. Some more recent examples are included .…”

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

“…The purities of all purification methods are statistically similar to the baseline-positive control from analysis by nonreduced CE-SDS, reduced CE-SDS, RP-HPLC, HP-SEC, HP-CEX, and N-Glycan. Sanaie et al reported a discrepancy between the resin-plate performance to that of a packed column in terms of glycoform distribution, but this was attributed to lower theoretical plates for the slurry method, which would not be a concern for affinity chromatography [26]. Variation is greater for the high-throughput formats than the baseline column format for some assays, but not all, indicating reasonable precision for the HTS formats relative to the baseline column format.…”