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Date Permissions Signed

6-4-2018

Date of Award

Spring 2018

Document Type

Masters Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

First Advisor

Spiegel, P. Clint

Second Advisor

Prody, Gerry

Third Advisor

Antos, John M.

Abstract

Blood coagulation factor VIII (FVIII) is a non-enzymatic protein cofactor, which plays a crucial role in the formation of a stable blood clot. Absence or deficiency of FVIII results in the blood disorder hemophilia A, with symptoms including internal hemorrhaging and the inability to stop bleeding from open wounds. Treatment of hemophilia A relies on replacement of FVIII with blood, plasma, or protein concentrate infusions. Unfortunately, approximately 30% of patients receiving replacement FVIII generate pathologic anti-FVIII inhibitory antibodies, which both reduce the effectiveness of the FVIII therapeutic and increase the severity of hemophilia A symptoms.

This thesis reports the determination of the molecular structure for “Et3i”, a next-generation human/porcine chimeric FVIII protein for hemophilia A therapy. At 3.2 Å resolution with a Rwork of 0.2146 and Rfree of 0.2879, this will be the highest resolution structure of FVIII to date and will be of substantial interest to the hematological community. Furthermore, an improved model of human FVIII with more robust geometry and amino acid register assignment, and a Rwork of 0.2655, and Rfree of 0.2895 based on previous 3.7 Å data has been constructed. Lastly, progress has been made towards the structural determination of the inhibitory antibodies M6143, 2A9, and B136 in complex with the C1 domain of human FVIII. Details of these interactions could inform the development of future hemophilia A protein therapeutics with reduced immunogenicity.

Type

Text

Keywords

X-ray Crystallography, Biochemistry, Hemophilia A, Blood Coagulation Factor VIII (FVIII), Structural Biology

DOI

https://doi.org/10.25710/wn94-rc05

Publisher

Western Washington University

OCLC Number

1039725210

Subject – LCSH

Hemophilia--Treatment--Complications; Hemophilia--Treatment--Research; Blood coagulation factor VIII--Therapeutic use--Testing; Blood coagulation factor VIII antibodies; Chimerism--Testing; Blood coagulation disorders; Therapeutics, Experimental

Format

application/pdf

Genre/Form

masters theses

Language

English

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 thesis for commercial purposes, or for financial gain, shall not be allowed without the author's written permission.

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