Biotech therapeutics such as antigen-binding fragments (Fab) have emerged as effective treatments for many hitherto untreatable modern diseases. However, affordability remains a daunting challenge for most of the world. Advancements in manufacturing, such as continuous manufacturing, have the potential to offer higher productivity and better economics without sacrificing product quality. In this paper, we propose an integrated continuous purification process for a Fab therapeutic (Ranibizumab). A batch purification process was initially developed followed by its conversion to continuous format. Enabling technologies like coiled flow inversion reactor, inline concentrator, concurrent chromatography, and membrane chromatography were utilized for the successful conversion of batch to continuous process. Suitable process modifications were made in order to ensure efficient conversion and integration of different unit operations. The final integrated continuous process consisted of continuous refolding and precipitation in a coiled flow inverter reactor, inline concentration using inline concentrator, multicolumn capture, multicolumn polishing concurrent chromatography, and an anion exchange membrane adsorber. The continuous bioprocessing train was run uninterrupted for 50 h to demonstrate its capability and the resulting output was analyzed for the various critical quality attributes. All attributes were found to be consistent over the period of operation with product purity &gt; 99 %, high molecular weight impurities &lt; 0.05 %, HCP &lt; 10 ppm and DNA &lt; 5ppb. The developed assembly offers higher productivity, fewer hold steps, and significantly higher equipment, and resin utilization. We hope that the current study will encourage continuous processing as a method to improve manufacturability of difficult to refold, low yield molecules like Fabs.

Integrated continuous purification for antigen-binding fragments (Fab) expressed as inclusion bodies: Ranibizumab as a case study

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.