This document summarizes information about factor VIII, which is involved in the intrinsic pathway of coagulation. It discusses that factor VIII is produced in liver sinusoidal cells and endothelial cells lining blood vessels. It consists of six domains and undergoes proteolytic processing. The document also describes the production and purification process of recombinant factor VIII from human embryonic kidney cells for use in treatments like Nuwiq, which has advantages over plasma-derived factor VIII.

2.  Name : Antihaemophilic factor
 Source: Liver sinusoidal cells and Endothelium lining blood
vessel and platelets(outside liver)
 Pathway: intrinsic
 Activator: Thrombin
 Action: work with factor IX and calcium to activate factor X
 Deficiency: Haemophilia A

4.  FVIII:C
• Smaller polypeptide:
170kDa
• Exihibit coagulant
activity
 FVIII:vWF
• Larger polypeptide,
Von Willebrand factor
• Associated with
Platelet adhesion
Factor VIII complex

5.  Factor VIII polypeptide consists of
2332 amino acid residues and
include six domains: A1-A2-B-A3-
C1-C2, and is homologous to factor
V.
 A domains are homologous to the
A domains of the copper-binding
protein ceruloplasmin.
 The C domains belong to the
phospholipid-binding discoidin
domain family.
 Three acidic subdomains:
A1(a1)-A2(a2)-B-(a3)A3-C1-C2, localized at
boundary of A domains

6.  Proteolytic processing of FVIII.
• Black triangles – sites of
processing by the PACE/furin
family proteases;
• green triangles – sites of
processing during activation;
• Red triangles – sites of
processing during inactivation;
and
• blue circle represents the
coordinated copper ion

8. Plasma
Sepharose 4 Fast Flow
Q Sepharose High Performance
Virus inactivation
SP Sepharose High Performance
Superose 12prep grade
Ultrafilteration
Sterile filteration
Lyophilization
Dry Heat Treatment
Factor VIII

10.  Cell line : Human embryonic kidney(HEK) 293F cells
 Advantage : Elimination of risk of accidental transmission of Blood-
borne diseases.
 Overall yield is approx. 50%
 Has human post translational modifications
 Devoid of antigenic non-human epitopes
 Example : Nuwiq (Octapharma AG, Switzerland)

11. Human pdFVIII (without B domain)
Nuwiq