Automation Is Key
Finally, there is the issue of reversible versus irreversible associations. Time, temperature, salt conditions, and sample dilution can all contribute to changing chromatographic profiles that can cause significant problems.
“In early formulation and analytical development work, we generally have limited sample, many candidates, multiple projects, and not enough time,” said Darryl Davis, principal scientist, pharmaceutical development & manufacturing sciences, Janssen Research & Development.
“Automation of the sample preparation and analysis techniques can be useful and provide unexpected benefits when used in an integrated development environment where cell-line selection, purification, and formulation have intersecting data points from the same sample.
“We have an automated high-throughput screening platform in place that can screen the formulation space to look for aggregation, turbidity, solubility, and product quality. The platform can use dynamic light scattering, differential scanning calorimetry, UV absorbance, and liquid chromatography/liquid chromatography-mass spectrometry (LC/LC-MS) assays to test critical quality attributes,” he explained.
“The cell biology group has used the LC-MS technique to screen for glycosylation and clipping of 96 well-plate transfectants. They have also used it to look at issues of scale and media changes. The amount of samples being run and analyzed would not be possible without using these automated platforms. The push toward standardization or platforming of techniques allows for streamlining the overall process and ensures a lower failure rate,” Davis remarked.
“We are looking at the fundamental mechanisms of folding and unfolding. When does a protein begin to unfold to form the first seeds, the first nuclei that will begin to aggregate?”