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Modified harvest system for enhancing Factor VIII yield in alternating tangential flow perfusion culture

Paper ID Volume ID Publish Year Pages File Format Full-Text
20118 43158 2016 5 PDF Available
Title
Modified harvest system for enhancing Factor VIII yield in alternating tangential flow perfusion culture
Abstract

•We developed a novel harvest system for ATF perfusion culture as an alternative method to replace a peristaltic pump.•The novel harvest system consists of a check valve, pinch valve and timer.•The system achieved the enhancement of the yield of Factor VIII activity and Factor VIII antigen.•The yield enhancement was due to the increase of transmembrane pressure across hollow fiber filter using the system.•The system offers the possibility to overcome the fouling problem in ATF perfusion culture.

This study describes the development and experimental verification of a modified harvest system to enhance Factor VIII (FVIII) yield in an alternating tangential flow (ATF) perfusion culture. The main innovation of the modified harvest system is the use of check and pinch valves, eliminating the need of a peristaltic pump for harvest. The system was applied to perfusion cultures of Chinese hamster ovary cells, which co-express both recombinant human FVIII (rhFVIII) and von Willebrand factor (vWF). The modified harvest system showed comparable cell growth with the conventional harvest system using a peristaltic pump. The perfusion rate was successfully controlled using the system. In addition, the modified harvest system achieved an approximately 13.6-fold increase in the final concentration yield of FVIII activity and a 1.47-fold increase in the production yield of FVIII activity compared with a peristaltic pump. Enhancement of the yield of FVIII activity resulted from the reduction of FVIII antigen (FVIII:Ag) retention. As a result of transmembrane pressure (TMP) measurement, the reduction of the retained FVIII:Ag was due to the increased TMP, which was caused by the characteristic function of a check valve, compared with a peristaltic harvest system. The modified harvest system developed in this study could be useful to enhance the production yield of other recombinant proteins in ATF perfusion culture.

Keywords
Modified harvest system; Alternating tangential flow perfusion culture; FVIII yield; Pinch valve; Check valve; Transmembrane pressure; Filter fouling
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Modified harvest system for enhancing Factor VIII yield in alternating tangential flow perfusion culture
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 121, Issue 5, May 2016, Pages 561–565
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us