Blood Clotting Factor VIII: From Evolution to Therapy

Орлова, Н. А.; Ковнир, С. В.; Vorobiev, I. I.; Gabibov, A. G.; Vorobiev, A. I.

doi:10.32607/20758251-2013-5-2-19-39

Cited by 51 publications

(62 citation statements)

References 187 publications

(249 reference statements)

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“…FVIII is a heterodimer stabilized in plasma by copper ions and via binding to VWF [38]. Loss of the copper ion leads to dissociation and loss of activity.…”

Section: Overview: Assessing the Quality Of Transfusable Plasmamentioning

confidence: 99%

Quality Assessment of Established and Emerging Blood Components for Transfusion

Acker

Marks

Sheffield

2016

Journal of Blood Transfusion

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Blood is donated either as whole blood, with subsequent component processing, or through the use of apheresis devices that extract one or more components and return the rest of the donation to the donor. Blood component therapy supplanted whole blood transfusion in industrialized countries in the middle of the twentieth century and remains the standard of care for the majority of patients receiving a transfusion. Traditionally, blood has been processed into three main blood products: red blood cell concentrates; platelet concentrates; and transfusable plasma. Ensuring that these products are of high quality and that they deliver their intended benefits to patients throughout their shelf-life is a complex task. Further complexity has been added with the development of products stored under nonstandard conditions or subjected to additional manufacturing steps (e.g., cryopreserved platelets, irradiated red cells, and lyophilized plasma). Here we review established and emerging methodologies for assessing blood product quality and address controversies and uncertainties in this thriving and active field of investigation.

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“…FVIII is a heterodimer stabilized in plasma by copper ions and via binding to VWF [38]. Loss of the copper ion leads to dissociation and loss of activity.…”

Section: Overview: Assessing the Quality Of Transfusable Plasmamentioning

confidence: 99%

Quality Assessment of Established and Emerging Blood Components for Transfusion

Acker

Marks

Sheffield

2016

Journal of Blood Transfusion

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“…In the case of a blood coagulation event, FVIII undergoes specific proteolytic cleavage by the thrombin, converting it to the activated FVIII (FVIIIa) [2]. FVIIIa dissociates from vWF, binds to the surface of acidic cell membranes of activated platelets, recruits binding of the activated factor IX (FIXa) and the factor X (FX) to the ternary complex “X-ase” (or tenase), promotes specific cleavage of the FX by the FIXa to the activated FX (FXa) and allows the release of the FXa from the X-ase complex [3]. Natural human FVIII is a large glycoprotein with multiple size variants, ranging from 190 to 300 kDa.…”

Section: Introductionmentioning

confidence: 99%

Stable high-level expression of factor VIII in Chinese hamster ovary cells in improved elongation factor-1 alpha-based system

et al. 2017

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BackgroundRecombinant factor VIII (FVIII), used for haemophilia A therapy, is one of the most challenging among the therapeutic proteins produced in heterologous expression systems. Deletion variant of FVIII, in which the entire domain B is replaced by a short linker peptide, was approved for medical use. Efficacy and safety of this FVIII deletion variant are similar to full-length FVIII preparations while the level of production in CHO cells is substantially higher.Typical levels of productivity for CHO cell lines producing deletion variant FVIII-BDD SQ, described elsewhere, are 0.5–2 IU/ml, corresponding to the concentration of FVIII of about 0.2 μg/ml. Using standard vectors based on the cytomegalovirus promoter (CMV) and the dihydrofolate reductase cDNA we have previously obtained the cell line secreting 0.5 IU/ml of FVIII-BDD, which roughly corresponds to the previously published data.ResultsAn expression system based on CHO genomic sequences including CHO-EEF1A promoter and Epstein-Barr virus terminal repeat fragment allowed us to achieve 80-fold increase in the production level as compared with the conventional expression system based on the CMV promoter.Immediately after the primary selection FVIII -producing cells secreted 5–10 IU/ml of FVIII-BDD, and after multi-stage methotrexate-driven amplification a stable clonal line 11A4H was selected, secreting 39 IU/ml of FVIII-BDD in the simple batch culturing conditions, which considerably exceeds known indicators for industrial producers of this protein. In contrast to other FVIII-BDD producing lines 11A4H accumulates low proportion of the secreted FVIII on the membrane. Its productivity may be further increased approximately two-fold by adding sodium butyrate and butylated hydroxyanisol to the culture medium.A five-stage purification process for the factor VIII was employed. It allowed isolation of the intact FVIII-BDD as was confirmed by mass spectrometry. Purified FVIII-BDD has a specific activity of 11,000 IU/mg, similar to known recombinant FVIII drugs.ConclusionsThe recombinant FVIII-BDD was produced in CHO cells without addition of any animal-derived materials, purified and characterized. Novel genetic constructions for the expression of heterologous proteins combined with optimized cultivation method allowed to obtain the secretion level of biologically active recombinant FVIII increased by almost ten times as compared with the previously published analogues.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-017-0353-6) contains supplementary material, which is available to authorized users.

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“…FVIII gene protein production results in the production of a single polypeptide that is modified and cleaved to form light and heavy chains. These light and heavy chains are held together by a labile metal ion bridge that is required for proper functioning of the protein 3. It was found that deleting most of the B domain of FVIII actually improved secretion from the cell in the recombinant process,12 and so B-domain-deleted (BDD) FVIII products such as ReFacto ® were developed.…”

Section: Current Factors Available and Extended Half-life (Ehl) Factomentioning

confidence: 99%

“…The cloning of the gene for FVIII in 1984 resulted in recombinant FVIII preparations becoming available in the 1990s. Currently, the standard of care in developed countries is to offer primary prophylactic FVIII infusions to patients with severe HA and has led to dramatic increase in the quality of life for these patients 2,3…”

Section: Introductionmentioning

confidence: 99%

Many factor VIII products available in the treatment of hemophilia A: an embarrassment of riches?

Lieuw¹

2017

JBM

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Hemophilia A (HA) is a common bleeding disorder caused by the deficiency of factor VIII (FVIII) with an incidence of ~1 in 5000 male births. Replacement of FVIII is necessary to prevent and treat bleeding episodes. However, with multiple new drugs in addition to old standards, choosing among the different FVIII treatment options is harder than ever. There are FVIII products that are plasma derived or recombinant, FVIII products designed to extend the half-life of FVIII, and the first single-chain FVIII product, recombinant factor VIII single chain (rFVIII-SC). As development of inhibitors to FVIII continues to be a major problem in the care of HA patients, recent studies showing lower rates of inhibitor development with plasma-derived FVIIII products versus recombinant FVIII products have made choosing among the many options now available even more complex. Although still unproven, extended half-life (EHL) products may provide the hope of decreased immunogenicity but need further testing in previously untreated patients (PUPs). This review highlights some of the differences between FVIII products currently available and hopefully assists the clinician to decide which FVIII product to choose for their patients.

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Blood Clotting Factor VIII: From Evolution to Therapy

Cited by 51 publications

References 187 publications

Quality Assessment of Established and Emerging Blood Components for Transfusion

Quality Assessment of Established and Emerging Blood Components for Transfusion

Stable high-level expression of factor VIII in Chinese hamster ovary cells in improved elongation factor-1 alpha-based system

Many factor VIII products available in the treatment of hemophilia A: an embarrassment of riches?

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