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Comparative Study
. 2010 Jun;8 Suppl 3(Suppl 3):s98-104.
doi: 10.2450/2010.016S.

Comparison Among Plasma-Derived Clotting Factor VIII by Using Monodimensional Gel Electrophoresis and Mass Spectrometry

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Free PMC article
Comparative Study

Comparison Among Plasma-Derived Clotting Factor VIII by Using Monodimensional Gel Electrophoresis and Mass Spectrometry

Anna Maria Timperio et al. Blood Transfus. .
Free PMC article

Abstract

Background: Deficiency or dysfunction of coagulation factor VIII (FVIII) is the underlying cause of haemophilia A. Haemophilic patients are at present treated with plasma-derived FVIII (pdFVIII) or recombinant FVIII (rFVIII) in order to correct their clotting deficiency. pdFVIII concentrates are exclusively produced from human plasma upon pooling from multiple donors. It is not know whether the presence of excess of other plasma proteins, in addition to von Willebrand factor, could stimulate untoward immune responses in the recipient. Thus, information regarding the presence of contaminants in commercial products is of concern.

Materials and methods: Two commercially available pdFVIII concentrates were characterized through SDS-PAGE and mass spectrometry Emoclot and Beriate.

Results: The components of two pdFVIII products considered in this study were well identified by mass spectrometry analysis, in both cases we found abundant components coming from blood plasma, and some other contaminants. Only in Beriate we also found truncated form of pdFVIII.

Conclusion: The two pdFVIII examined showed the presence of vWF, Fibrinogen in excess, and other substances that could be considered as contaminants or impurities.

Keywords: SDS-PAGE; contaminants; haemophilia A; mass spectrometry; plasma derived FVIII.

Figures

Figure 1
Figure 1
Mini gel SDS PAGE of two pdVIII preparations: Emoclot ® 1000 UI/10 mL batch KA0805 Kedrion and Beriate® P1000 batch 81865011E CLS Behring. Samples are reduced according to Laemmli’s method and are analysed at different concentration of acrilamide a) 7,5% b) 9% c) 11% d) 12%.
Figure 2
Figure 2
SDS-PAGE (7.5%) of reduced samples (pdFVIII) according to Laemmli’s method. Protein are detected by Coomassie Brilliant Blue. Lane 1 HMW standard, Lane 2: 40 ug of pdFVIII Emoclot ® 1000 UI/10 mL batch KA0805 Kedrion, Lane 3: 40 ug of pdFVIII Beriate® P1000 batch 81865011E CLS Behring.

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