The biopharmaceutical industry is strongly moving to high-throughput formats for process development. Powerful analytics are required as an essential and integral part of this approach and must deliver short analysis time, appropriate resolution, efficient data handling, and flexible output options. Evolving from an 80 minutes routine protocol, we have established a Reversed phase (RP) HPLC method with a run time of only 8 minutes as a rapid in-process analytical tool to support high-throughput insulin purification development, where 100 to 300 samples per day may be generated from resin screening trials. The short method, which uses a Chromolith® HighResolution RP-18 endcapped 100-4.6 column, maintains adequate selectivity in the relevant region of separation of the target insulin form and closely related impurities, such as the des-amido insulin molecule. We will demonstrate the comparability of both methods with purity and titer data from a range of process samples. With a second focus on method reliability, we have assessed and optimised the evaluation of analytical chromatograms. Blank subtraction, peak integration methods, peak integration window, and linearity of peak signals were all identified as crucial for accuracy and reproducibility.

Rapid and reliable analytical RP HPLC for high-throughput insulin process development

253rd National Meeting (2017)

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