Synthesis of recombinant blood coagulation factor VIII (FVIII) heavy and light chains and reconstitution of active form of FVIII

Oh, Seoung Wug; Lee, Mi Young; Song, Dong Weon

doi:10.1038/emm.1999.16

Cited by 4 publications

(3 citation statements)

References 20 publications

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“…During the last 6 years, many new recombinant FVIII (rFVIII) and FIX (rFIX) concentrates have attracted the attention of treaters and patients due to their innovative features. Improvements of the manufacturing procedures of such concentrates, as for example, in the adoption of cultures of human embryonic kidney (HEK) cells, 1,2 more selective immunoaffinity chromatography, 3,4 co-expression of the albumin gene 5–7 or fragment crystallizable (Fc) region of immunoglobulin in HEK cell lines, 8–9 molecule modification 10–12 and/or PEGylation, 13–18 have increased the bioavailability of the rFIX concentrates and, to a lesser extent, rFVIII ones. 19 In this context, pharmacokinetics (PK), pharmacodynamics (PD) and pharmacoeconomics (PE) are the tools, which allow the treaters the evaluation of the patients’ unmet needs and the way they can be addressed by new CFCs.…”

Section: Introductionmentioning

confidence: 99%

Pharmacokinetic-based prediction of real-life dosing of extended half-life clotting factor concentrates on hemophilia

Morfini¹,

Gherardini

2018

Therapeutic Advances in Hematology

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The improvement of clotting factor concentrates (CFCs) has undergone an impressive boost during the last six years. Since 2010, several new recombinant factor (rF)VIII/IX concentrates entered phase I/II/III clinical trials. The improvements are related to the culture of human embryonic kidney (HEK) cells, post-translational glycosylation, PEGylation, and co-expression of the fragment crystallizable (Fc) region of immunoglobulin (Ig)G1 or albumin genes in the manufacturing procedures. The extended half-life (EHL) CFCs allow an increase of the interval between bolus administrations during prophylaxis, a very important advantage for patients with difficulties in venous access. Although the inhibitor risk has not been fully established, phase III studies have provided standard prophylaxis protocols, which, compared with on-demand treatment, have achieved very low annualized bleeding rates (ABRs). The key pharmacokinetics (PK) parameter to tailor patient therapy is clearance, which is more reliable than the half-life of CFCs; the clearance considers the decay rate of the drug concentration-time profile, while the half-life considers only the half concentration of the drug at a given time. To tailor the prophylaxis of hemophilia patients in real-life, we propose two formulae (expressed in terms of the clearance, trough and dose interval between prophylaxis), respectively based on the one- and two-compartmental models (CMs), for the prediction of the optimal single dose of EHL CFCs. Once the data from the time decay of the CFCs are fitted by the one- or two-CMs after an individual PK analysis, such formulae provide to the treater the optimal trade-off among trough and time-intervals between boluses. In this way, a sufficiently long time-interval between bolus administration could be guaranteed for a wider class of patients, with a preassigned level of the trough. Finally, a PK approach using repeated dosing is discussed, and some examples with new EHL CFCs are shown.

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Section: Introductionmentioning

confidence: 99%

Pharmacokinetic-based prediction of real-life dosing of extended half-life clotting factor concentrates on hemophilia

Morfini¹,

Gherardini

2018

Therapeutic Advances in Hematology

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“…Production of biologically active rFVIII through eukaryotic cell culture system has been demonstrated by several investigators (Woods et al, 1984;Kauffman et al, 1988), however, problems such as low productivity, inhibiting antibody formation and instability of the products still remain. In the previous study, we prepared a mutant cDNA of FVIII heavy chain by site-directed mutagenesis (G 1074 A 1074 ) and substitution of Arg 336 with Gln 336 for the production of rFVIII with high stability (Oh et al, 1999). In the present study, the mutant B-domain deleted rFVIII m expressed in baculovirus-insect cell (Sf9) was purified and its functional characteristics were determined.…”

Section: Introductionmentioning

confidence: 99%

“…In the present study, the mutant B-domain deleted rFVIII m expressed in baculovirus-insect cell (Sf9) was purified and its functional characteristics were determined. (Oh et al, 1999). FVIII-B m cDNA was inserted into pAcSG2 baculoviral transfer vector ( Figure 1) by subcloning the the cDNA into XhoI and NotI restriction enzyme recognizing sites of pAcSG2 according to the method described by Sambrook et al (1989).…”

Section: Introductionmentioning

confidence: 99%

Expression and characterization of a mutant recombinant blood coagulation factor VIII (rFVIII (m))

Cho²,

Jin³

et al. 2002

Exp Mol Med

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In an earlier study, a site directed mutant rFVIII (rFVIII m and native FVIII with APC decreased their cofactor activities, however, the former exhibited a slower decrease than the latter, although no significant difference was present. rFVIII m formed a complex with vWF, resulting in a stabilized form, and the lag period of thrombin-mediated activating was extended by vWF association. These results implicated that rFVIII m expressed in baculovirus-insect cell system had a comparable capacity as FVIII cofactor activity and might be a good candidate for the FVIII replacement therapy for hemophilia A patients.

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TMAO to the rescue of pathogenic protein variants

Kumari

Singh²,

Singh³

et al. 2022

Biochimica et Biophysica Acta (BBA) - General Subjects

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Synthesis of recombinant blood coagulation factor VIII (FVIII) heavy and light chains and reconstitution of active form of FVIII

Cited by 4 publications

References 20 publications

Pharmacokinetic-based prediction of real-life dosing of extended half-life clotting factor concentrates on hemophilia

Pharmacokinetic-based prediction of real-life dosing of extended half-life clotting factor concentrates on hemophilia

Expression and characterization of a mutant recombinant blood coagulation factor VIII (rFVIII (m))

TMAO to the rescue of pathogenic protein variants

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