Method Development

PhyNexus offers Advantages of Microscale, Direct Correlation to Large Scale

PhyNexus is forever changing the way method development and optimization is done.  With PhyNexus PhyTip Columns, analytical scale chromatography results directly correlate with preparative and manufacturing scale chromatography.  That means dozens, even hundreds of multi-variate, multi-parallel experiments at the miniature scale can be done in days to identify the very best candidates on the basis of yield, purity and activity (not just one parameter).  And the results will scale.  The benefits in terms of time, labor, cost and results prior to moving to lab scale operations are huge.

The Challenge to Accelerate Process Development

Method development and optimization techniques and technology are not keeping pace with advances in manufacturing scale production.  Lab scale method development is slow, often using large 10 mL columns and requiring large volumes of cell lysate.  The slow pace of lab scale purifications means the number of experiments is limited and focus is primarily on protein mass capture (yield) and, to a lesser extent, protein purity.  Traditional lab processes are not designed for efficient multivariate, multi-parallel experimentation for simultaneous optimization yield, purity and protein activity – and especially not done in days.

PhyTip Columns Accelerate & Improve Method Development

PhyNexus technology delivers an automated solution for process development and optimization at the miniature scale that correlates virtually directly to lab scale, preparative scale and even manufacturing scale chromatography.  The key is in the interaction of the PhyTip Column with the gentle back and forth flow of Dual Flow Chromatography, which drives the mobile / solid interaction to equilibrium much like the continuous loading of a laboratory column would do.

PhyTip Columns are compatible with most 8-, 12- and 96-channel robotic instruments, allowing for rapid and repeatable experimentation.  The small 1 mL size means only moderate sample volumes are required for many, many experiments.  And process time itself is vastly reduced.  Significant  benefits accrue from miniaturization, including fully automated multivariate, multi-parallel testing of various conditions on multi-channel robotics, significantly faster processing times, and analysis of the highest quality proteins selected for yield, purity and activity (including agglomeration prevalence).

Automating Process Development with PhyTip Columns

Merck & Co. pharmaceutical researchers used PhyNexus PhyTip Columns to miniaturize and automate the multivariate analysis necessary to optimize the fermentation conditions for the production of a recombinant vaccine for human papillomavirus (HPV).  In conclusion, the researchers found:

“Overall, the automated purification [using PhyNexus PhyTip Columns on a Tecan Freedom EVO 8-channel workstation] was concordant with the laboratory-scale method in the assessment of VLP [virus-like particles] recovery and purity, while improving throughput by over 10-fold and vastly reducing the labour of the purification…  The miniaturization and automation of the chromatographic purification have significantly decreased the time and labour required to evaluate yeast fermentations, while only requiring milligram amounts of cell input.”

Wenger, M. D., et al., An automated microscale chromatographic purification of virus-like particles as a strategy for process development. Biotechnol. Appl. Biochem. 2007, 47, 131-139.