Novel Attempt at Expression of Properly Folded C1, C1C2, and A2 Factor VIII Protein Domains in E. coli cells
Research Mentor(s)
Spiegel, P. Clint
Description
Hemophilia A is a bleeding disorder treated with synthetic blood coagulation factor VIII (fVIII). In order to study fVIII we currently must use protein grown exclusively in human cell lines - a process requireing bio-safety clearance II which can be prohibitively expensive. Previous attempts to express and purify domains of fVIII in prokaryotic cells have resulted in improperly folded protein eluting in the insoluble fraction, rendering it unusable. Here we covalently bind thioredoxine to the A2, C1C2, and C1 domains of the fVIII protein and grow them in SHuffle T7 express cell line in an attempt to keep the protein properly folded and soluble.
Document Type
Event
Start Date
14-5-2015 10:00 AM
End Date
14-5-2015 2:00 PM
Department
Chemistry
Genre/Form
student projects; posters
Subjects – Topical (LCSH)
Blood coagulation factor VIII; Blood proteins--Structure; Hemophilia--Treatment
Type
Image
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this documentation for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.
Language
English
Format
application/pdf
Novel Attempt at Expression of Properly Folded C1, C1C2, and A2 Factor VIII Protein Domains in E. coli cells
Hemophilia A is a bleeding disorder treated with synthetic blood coagulation factor VIII (fVIII). In order to study fVIII we currently must use protein grown exclusively in human cell lines - a process requireing bio-safety clearance II which can be prohibitively expensive. Previous attempts to express and purify domains of fVIII in prokaryotic cells have resulted in improperly folded protein eluting in the insoluble fraction, rendering it unusable. Here we covalently bind thioredoxine to the A2, C1C2, and C1 domains of the fVIII protein and grow them in SHuffle T7 express cell line in an attempt to keep the protein properly folded and soluble.