N. O. Shurko, V. L. Novak


The article deals with basic methods used by modern technology to obtain coagulation factor VIII (FVIII).
The blood plasma fractionation remains the only biotechnological approach to make life-saving protein therapy to treat human diseases. The biological medicines from human plasma play a vital role in the treatment of patients with different diseases. These products include a range of coagulation factors (FVIII, FIX, the prothrombin complex, Von Willebrand factor, fibrinogen etc.), immunoglobulins, protease inhibitors, anticoagulants and albumin.
Four plasma proteins are commercially important for production: albumin, IgG, factor VIII, and factor IX. VIII is a coagulation factor in the blood, which is missing or defective in patients with Hemophilia A. Replacement therapy with FVIII concentrates constitutes the basis for hemophilia care.
Cryoprecipitate was described in the mid 60's of the XX century as a first concentrate of antihemophilic FVIII.The main indications for the clinical use of cryoprecipitate were hypofibrinogenemia or disfibrinogenemia. Previously, cryoprecipitate was used for treatment of hemophilia A and von Willebrand’s disease.
Traditional FVIII production methods included deposition steps, which were aimed at elimination of protein impurities such as fibrinogen, fibronectin and immunoglobulins. These technologies could use the combination of methods at low temperatures or the addition of protein precipitating substances (PEG, polyvinylpyrrolidone, dextran, ficol, percol etc.). Using chromatographic methods in FVIII production technology allowed receiving high purity and specific activity concentrate of FVIII. Ion exchange chromatography techniques are often used in order to isolate coagulation FVIII. These techniques include methods of affinity chromatography as well as the use of monoclonal antibodies to bind of FVIII.
Nowadays, production of plasma concentrate of FVIII is used in combination with different chromatographic techniques.


coagulation; plasma; factor VIII (FVIII;, separation


Sravnenie dvuh alternativnyih metodov hromatograficheskoy ochistki pri poluchenii prepaarta VIII faktora svertyivaniya plazmyi krovi cheloveka v dozirovke 500 edinits/flakon / A. V. Yamkin, O. V. Stronin, L. N. Nikitina, N. A. Simenova, A. A. Epanchentseva. Sibirskiy meditsinskiy zhurnal (Tomsk). 2011. № 2–2, (26). S. 49–55. [in Russian]

Aldington S., Bonnerjea J. Scale-Up of monoclonal antibody purification processes. J. Chromatogr B Analyt Technol Biomed Life Sci. 2007. 848 (1). P. 64–78. doi: 10.1016/j.jchromb.2006.11.032.

BIVV001, a new class of factor VIII replacement for hemophilia A that is independent of von Willebrand factor in primates and mice / Chhabra E. S., Liu T., Kulman J., S. Patarroyo-White, B. Yang, Q. Lu et all. Blood. 2020. 135. (17). P.1484–1496.

Burnouf T., Burnouf-Radosevich M., Huart J. J., Goudemand M. A highly purified factor VIII:C concentrate pre-pared from cryoprecipitate by ion-exchange chromatog-raphy. Vox Sang. 1991. Vol. 60. (S1). P. 8–15. 3–15. doi:10.1111/j.1423-0410.1991.tb00864.x

Burnouf Т. New developments in the production of plasma derivatives. 2016., London. 20.

Development and validation of an affinity chromatography step using a peptide ligand for cGMP production of factor VIII / B. D. Kelley, M. Tannatt, R. Magnusson, S. Hagelberg. J. Booth. Biotechnol Bioeng. 2004. 5. 87(3). P. 400–412. doi:10.1002/bit.20124.

Development of a peptidomimetic ligand for efficient isolation and purification of factor VIII via affinity chromatography / S. Knor, A. V. Khrenov, B. Laufer, E. L. Saenko, C. A. Hauser, H. Kessler. J. Med. Chem. 2007. 50. (18). P. 4329–4339. doi:10.1021/jm070304x.

Federici A. B. Highly purified VWF/FVIII concentrates in the treatment and prophylaxis of the von Willebrand disease: the PRO.WILL Study. Haemophilia. 2007. 13 (5). Р.15–24. doi:10.1111/j.1365-2516.2007.01573.x.

Fuare A., Caron M., Tepenier D. Improved buffer for the chromatographic separation of Factor VIII coagulant. Journal of Chromatography. 1983. 257. P. 387–391. doi:10.1016/S0021-9673(01)88196-6.

Harrison P., Saundry R. H., Savidge G. F. Chromatography of the VIII/vWF complex on dextran sulphate sepharose. Thromb Res. 1988. 50. P. 295–304. doi: 10.1016/0049-3848(88)90230-7.

Labrou N. E. Design and selection of ligands for affinity chromatography. J. Chromatogr B Analyt Technol Biomed Life Sci. 2003. 25. 790 (1–2). P. 67–78. doi:10.1016/s1570-0232(03)00098-9.

Lack of recombinant factor VIII B-domain induces phospholipid vesicle aggregation: implications for the immunogenicity of factor VIII / K. Grushin, J. Miller, D. Dalm, E. T. Parker, J. E. Healey, P. Lollar, S. Stoilova-McPhie. Haemophilia. 2014. 20 (5). P. 723–731. doi:10.1111/hae.12421.

Large-scale purification of factor VIII by affinity chromatography: optimization of process parameters / P. W. M. Te Booy, A. Faber, E. De Jonge, E. P. Wolterink, W. Riethorst, T. Beugeling, A. B. J. Over, B. W.Köng. J Chromatogr. 1990. 9. 503 (1). P. 103–114. doi: 10.1016/S0021-9673(01)81494-1.

Mazurkiewicz-Pisarek A., Płucienniczak G., Ciach T., Płucienniczak A. The factor VIII protein and its function. Acta Biologica polonica. 2016. 63 (1). P. 11–16. doi: 10.18388/abp.2015_1056.

Morfini M., Coppola A., Franchini M., Di Minno G. Clinical use of factor VIII and factor IX concentrates. Blood Transfus. 2013. 11 (S.4). P. 55–63. doi: 10.2450/2013.011s.

Pat. US 20050239171 A1. Method for purifying factor VIII/vWF-complex by means of cation exchange chromatography // Mitterer A., Fischer B., Schonberger O., Thomas-Urban K., Dorner F., Eibl J. - Appl. 10/789,562 ; Fild 27.02.2004; Publ 27.10.2005.

Pat. US 5371195 A Method for purifying factor VIII and preparations obtained // Grandgeorge M., Lutsch C. Pasteur Merieux. – Appl. 07/948,395; Fild 23.09.1992; Publ 06.12.1994.

Pat. US 6605222 B1 Method for producing a factor VIII/von Willebrand factor complex / Linnau Y., Schoenhofer W. Baxter Aktiengesellschaft. – Appl. 09/623,245; Fild 25.02.1999; Publ 12.08.2003.

Pat. WO2009030866, C07K 1/ 18(2006.01), C07K 14/55(2006.01) Method for purifying the factor VIII and the von Willebrand factor / Poulle M., Bonneel P. LFB-Biotechnologies [FR/FR] – Appl. WO/2009/030866; Fild 28.08.2008; Publ 12.03.2009.

Progress in large-scale purification of factor VIII/von Willebrandfactor concentrates using ion-exchange chromatography / F. Mori, I. Nardini, P. Rossi, C. Nardini, C. Farina. Vox Sang. 2008. 95. P. 298–307. doi:10.1111/j.1423-0410.2008.01096.x

Purification of coagulation factor VIII by immobilized metal affinity chromatography / E. S. Rodrigues, C. L. Verinaud, D. S. Oliveira, I. Raw, A. P. Lopes, E. A. Martins, E. Cheng. Biotechnology and Applied Biochemistry. 2014. 62 (3). P. 343–348. doi: 10.1002/bab.1276.

Rotblat F., O’Brien D. P., O’Brien F. J. Purification of human factor VIII:C and its characterization by Western blotting using monoclonal antibodies. Biochemistry. 1985. 24. P. 4294–4300. doi: 10.1021/bi00337a007.

Te Booy M. P., Riethorst W., Faber A. Affinity purification of plasma proteins: characterization of six affinity matrices and their application for the isolation of human factor VIII. Thromb Haemost. 1989. 61. P. 234–237.



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