PB1106 - A novel and versatile platform for high expression of different FVIII modalities

Tuesday, June 25, 2024

13:45 – 14:45 ICT

Room: Exhibition Hall

Co-Author(s)

XL

Xu Liu

CMC Associate Director
Neoletix Biotech
Beijing, Beijing, China (People's Republic)
YL

Yang-li Lu

Sichuan Yuanda Shuyang Pharmaceutical Co., Ltd.
Chengdu, Sichuan, China (People's Republic)

Co-Author(s)

YZ

Yi Zhou

Sichuan Yuanda Shuyang Pharmaceutical Co., Ltd.
Chengdu, Sichuan, China (People's Republic)

Co-Author(s)

zW

zhijun Wang

AD Associate Director
Neoletix Biotech
Beijing, Beijing, China (People's Republic)

Co-Author(s)

XW

Xiao-zhou Wang

Sichuan Yuanda Shuyang Pharmaceutical Co., Ltd.
Chengdu, Sichuan, China (People's Republic)

Co-Author(s)

QL

Qianhui Li

R&D Manager
Neoletix Biotech
Beijing, Beijing, China (People's Republic)
YH

Yu He

Sichuan Yuanda Shuyang Pharmaceutical Co., Ltd.
Chengdu, Sichuan, China (People's Republic)
LW

Lin-feng Wu

Sichuan Yuanda Shuyang Pharmaceutical Co., Ltd.
Chengdu, Sichuan, China (People's Republic)

Presenting Author(s)

QL

Qi Li

Director
Neoletix Biotech
Lawrenceville, Georgia, United States

Background: Factor VIII (FVIII) replacement therapy continues to represent the current preferred modality for treatment of hemophilia A patients. Unfortunately, the difficulty in producing large amounts of high-quality FVIII translates to a high cost of goods and expensive final drug product. This is partly due to the low expression of FVIII and the need to compensate for this by using large bioreactors and/or continuous perfusion. We sought to evaluate the possibility to increase production of various FVIII modalities by utilizing strategies involving its binding chaperone, von Willebrand factor (vWF).

Aims: To determine if modified variants of vWF could be used to support greater expression of different FVIII molecules, increase productivity and substantially reduce cost.

Methods: A variety of truncated and/or mutated and modified vWF domains were tested for binding to FVIII and providing its stabilization in culture media. Modified vWF was found to provide increased stability and straightforward purification of a few different kinds of FVIIIs. These forms were further tested by a variety of functional methods to ensure comparability to other commercial FVIII molecules, especially when expressed at high levels.

Results: We have found that co-expression of various FVIII molecules (i.e., human, porcine, long-acting) with mutated and Fc-modified von Willebrand factor (vWF) can be produced at bioreactor expression levels (1000-6000 IU/ml) that significantly exceed current commercial productivities. Purification of the expressed FVIIIs is simplified by an initial Protein A purification step. FVIIIs prepared by this expression platform were biochemically characterized and their molecular interactions with other coagulation factors assessed.

Conclusion(s): Co-expression of modified vWF has been used to significantly increase FVIII expression in commercial production. Both human and porcine FVIII have been produced in large quantities and shown to have biochemical properties as well pharmacokinetic and pharmacodynamic attributes expected for commercially-produced FVIII.