Using high throughput screening to define virus clearance by chromatography resins.

Area of Science:

• Biotechnology
• Process Development
• Downstream Processing

Background:

• Chromatography resins are crucial for downstream process development, aiding in the removal of impurities and viral contaminants.
• Rodent cell lines, commonly used in biopharmaceutical production, can express retrovirus-like particles, necessitating effective clearance strategies.

Purpose of the Study:

• To evaluate the feasibility of using high throughput screening (HTS) in a 96-well batch-binding format for studying the removal of xenotropic murine leukemia virus (xMuLV).
• To compare the performance of an anion exchange resin (Q Sepharose Fast Flow™) and a mixed-mode resin (Capto adhere™) for viral clearance using HTS.

Main Methods:

• Developed and utilized a 96-well batch-binding HTS assay.
• Investigated the binding of xMuLV to Q Sepharose Fast Flow™ and Capto adhere™ under varying pH, NaCl concentrations, and impurity levels.
• Compared HTS data with traditional column chromatography results.

Main Results:

• HTS batch-binding data showed good agreement with column format results for virus retention.
• NaCl concentration and impurity level were identified as key parameters affecting xMuLV binding to both resins, while pH had minimal impact.
• Capto adhere™ demonstrated higher tolerance to NaCl and impurities compared to Q Sepharose Fast Flow™, with some product-specific binding effects observed.

Conclusions:

• The 96-well batch-binding HTS technique is an effective tool for rapidly defining conditions for robust virus clearance on chromatography resins.
• This HTS approach accelerates downstream process development by providing rapid insights into resin performance for viral clearance.
• Understanding the impact of process parameters like salt concentration and impurity levels is critical for optimizing viral clearance strategies.