Adaptation to high throughput batch chromatography enhances multivariate screening

Area of Science:

• Biotechnology
• Chemical Engineering
• Biopharmaceutical Development

Background:

• High-throughput process development enables exploration of complex design spaces with reduced material consumption.
• Batch chromatography in 96-well plates is typically used for screening buffer conditions or resins, not protein loading.
• Existing methods limit the multivariate analysis of chromatographic process parameters.

Purpose of the Study:

• To introduce a modified batch chromatography approach for expanded multivariate screening.
• To enable simultaneous assessment of protein loading and other buffer conditions.
• To enhance statistical power and reduce follow-up experiments in process development.

Main Methods:

• Development of the iterative batch chromatography (IBC) approach.
• Utilizing a liquid handling system for staggered loading in a 96-well plate format.
• Treating each well as an individual experiment to vary multiple parameters.

Main Results:

• The IBC approach allows for exploration of a larger multivariate experimental space.
• Characterization of interactions between protein loading, wash, and elution buffer conditions.
• Demonstration of the IBC method's utility with two specific examples.

Conclusions:

• Iterative batch chromatography significantly improves throughput and statistical power in process development.
• The IBC method facilitates comprehensive assessment of critical process parameters, including protein loading.
• This approach reduces the need for subsequent confirmatory experiments, accelerating bioprocess optimization.