Blood Research  2015;50(2):103-108

doi:10.5045/br.2015.50.2.103

Human coagulation factor VIII domain-specific recombinant polypeptide expression.

Su Jin CHOI 1 ; Ki Jung JANG ; Jeong A LIM ; Hye Sun KIM

Affiliations

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Keywords

Hemophilia A; Coagulation factor VIII; Coagulation factor IX; Domain-specific recombinant FVIII; Hep3B hepatocytes

Country

Republic of Korea

Language

English

Abstract

BACKGROUND: Hemophilia A is caused by heterogeneous mutations in F8. Coagulation factor VIII (FVIII), the product of F8, is composed of multiple domains designated A1-A2-B-A3-C1-C2. FVIII is known to interact with diverse proteins, and this characteristic may be important for hemostasis. However, little is known about domain-specific functions or their specific binding partners. METHODS: To determine F8 domain-specific functions during blood coagulation, the FVIII domains A1, A2, A3, and C were cloned from Hep3B hepatocytes. Domain-specific recombinant polypeptides were glutathione S-transferase (GST)- or polyhistidine (His)-tagged, over-expressed in bacteria, and purified by specific affinity chromatography. RESULTS: Recombinant polypeptides of predicted sizes were obtained. The GST-tagged A2 polypeptide interacted with coagulation factor IX, which is known to bind the A2 domain of activated FVIII. CONCLUSION: Recombinant, domain-specific polypeptides are useful tools to study the domain-specific functions of FVIII during the coagulation process, and they may be used for production of domain-specific antibodies.