Characterisation of the post-translational modifications of a novel, human cell line-derived recombinant human factor VIII

Kannicht, Christoph; Ramström, Margareta; Kohla, Guido; Tiemeyer, Maya; Casademunt, Elisabeth; Walter, Olaf; Sandberg, Helena

doi:10.1016/j.thromres.2012.09.011

Cited by 108 publications

(177 citation statements)

References 60 publications

Supporting

Mentioning

163

Contrasting

Unclassified

Order By: Relevance

“…Nielsen and colleagues examined extracted ion chromatograms showing sulfated and nonsulfated (minor or absent peak) Tyr1680-containing peptides. 59 Plasma-derived FVIII and recombinant FVIII (rFVIII) from human HEK293 cells demonstrated only a sulfated peak, 60 whereas rFVIII expressed by rodent cells exhibited both a sulfated peak and nonsulfated peaks. Likewise, a B-domain-deleted rFVIII expressed in Chinese hamster ovary (CHO) cells showed full Tyr1680 sulfation, suggesting that culture conditions may influence the efficiency of this modification.…”

Section: Fviii and Vwf Structural Biologymentioning

confidence: 99%

Life in the shadow of a dominant partner: the FVIII-VWF association and its clinical implications for hemophilia A

Pipe

Montgomery

Pratt

et al. 2016

Blood

184

186

View full text Add to dashboard Cite

A normal hemostatic response to vascular injury requires both factor VIII (FVIII) and von Willebrand factor (VWF). In plasma, VWF and FVIII normally circulate as a noncovalent complex, and each has a critical function in the maintenance of hemostasis. Furthermore, the interaction between VWF and FVIII plays a crucial role in FVIII function, immunogenicity, and clearance, with VWF essentially serving as a chaperone for FVIII. Several novel recombinant FVIII (rFVIII) therapies for hemophilia A have been in clinical development, which aim to increase the half-life of FVIII (∼12 hours) and reduce dosing frequency by utilizing bioengineering techniques including PEGylation, Fc fusion, and single-chain design. However, these approaches have achieved only moderate increases in halflife of 1.5-to 2-fold compared with marketed FVIII products. Clearance of PEGylated rFVIII, rFVIIIFc, and rVIII-SingleChain is still regulated to a large extent by interaction with VWF. Therefore, the half-life of VWF (∼15 hours) appears to be the limiting factor that has confounded attempts to extend the half-life of rFVIII. A greater understanding of the interaction between FVIII and VWF is required to drive novel bioengineering strategies for products that either prolong the survival of VWF or limit VWF-mediated clearance of FVIII. (Blood.

show abstract

Section: Fviii and Vwf Structural Biologymentioning

confidence: 99%

Life in the shadow of a dominant partner: the FVIII-VWF association and its clinical implications for hemophilia A

Pipe

Montgomery

Pratt

et al. 2016

Blood

184

186

View full text Add to dashboard Cite

show abstract

“…[48][49][50][51][52] In addition, certain rFVIII products may appear with poor sulfation at tyrosine 1680 (Tyr1680). 48 Interestingly, Tyr1680 sulfation is indispensable to the interaction between FVIII and its natural chaperone protein, VWF. 48,53,54 However, at present, there is no scientific evidence that differences in glycosylation and sulfation patterns between pdFVIII and rFVIII are of relevance to FVIII immunogenicity.…”

Section: Non-human Posttranslational Modificationsmentioning

confidence: 99%

“…48,49 All rFVIII products marketed so far are produced in Chinese hamster ovary (CHO) or baby hamster kidney (BHK) cells. These cells generate rodent-type glycosylation and sulfation patterns, which might enhance FVIII immunogenicity in several ways.…”

Section: Non-human Posttranslational Modificationsmentioning

confidence: 99%

“…Gal-alpha 1-3-Gal (alpha-Gal) and N-glycolylneuraminic acid (Neu5Gc) were detected in great abundance on rFVIII, which, in other contexts than hemophilia A, are known to elicit a marked immune response in all humans. [48][49][50][51][52] In addition, certain rFVIII products may appear with poor sulfation at tyrosine 1680 (Tyr1680). 48 Interestingly, Tyr1680 sulfation is indispensable to the interaction between FVIII and its natural chaperone protein, VWF.…”

Section: Non-human Posttranslational Modificationsmentioning

confidence: 99%

See 1 more Smart Citation

Alloantibodies to therapeutic factor VIII in hemophilia A: the role of von Willebrand factor in regulating factor VIII immunogenicity

Oldenburg¹,

Lacroix‐Desmazes

Lillicrap

2015

Haematologica

View full text Add to dashboard Cite

The rising incidence of neutralizing antibodies (inhibitors) against therapeutic factor VIII prompted the conduct of studies to answer the question as to whether this rise is related to the introduction of recombinant factor VIII products. The present article summarizes current opinions and results of non-clinical and clinical studies on the immunogenic potential of recombinant compared to plasma-derived factor VIII concentrates. Numerous studies provided circumstantial evidence that von Willebrand factor, the natural chaperone protein present in plasma-derived factor VIII products, plays an important role in protecting exogenous factor VIII from uptake by antigen presenting cells and from recognition by immune effectors. However, the definite contribution of von Willebrand factor in reducing the inhibitor risk and in the achievement of immune tolerance is still under debate. ABSTRACTinhibitors in stable PTPs after switching treatment is consistently lower than in previously untreated patients (PUPs), 31,32 probably because many of them have developed some kind of cross-tolerance. Thus, the immunogenicity of a given FVIII concentrate in PTPs is relevant for PTPs only. However, in daily clinical practice, more situations that demand a decision between treatment options are related to PUPs, i.e. patients in their early years of life who are newly diagnosed with hemophilia A or who sustain their first bleeding episode. 27,33 Are inhibitor testing methods a critical confounder?One of the most cited arguments used to confute a higher incidence of inhibitors associated with rFVIII treatment is that the testing frequencies and methods used for the detection of inhibitors have increased and improved over time. These improvements coincide with the introduction of rFVIII, which might have favored the detection of borderline and transient inhibitors in rFVIII-treated patient groups. 23,31,34 In fact, in 1995, the so-called "Nijmegen" method, a modification of the till then most widely used "Bethesda assay", was launched. 35 However, despite the improvements made over recent years, the methods used to detect inhibitors have not yet been standardized, 28,36 and the classical Bethesda assay originally published in 1975 is still the most frequently used assay. 34,36 Furthermore, the major advantage of the Nijmegen modification of the Bethesda assay lies in the improvement in the test's specificity near the cut-off value. 28,35,37 Hence, the advent and use of the Nijmegen assay should have caused a decrease in the rate of false positive test results and, consequently, a decrease rather than an increase in the inhibitor incidence in studies dating back to the 1990s. On the other hand, it is very possible that the increased testing frequency in the last two decades had an impact on the reported incidence of inhibitors. Studies reporting on the incidence of high-responding inhibitors [> 5 Bethesda units (BU) per milliliter] (Figure 1) avoided potential bias from assay performance and testing frequencies, because high-res...

show abstract

“…24,41 Although the progress in the development of products and several immune tolerance induction (ITI) studies that have improved inhibitor management, it remains the major issue in the hemophilia treatment considering both the patient's perspective as well as clinician's and healthcare provider's perspective. 42 In a recent publication, reported the results of a randomized trial to assess the incidence of FVIII inhibitors among patients treated with plasma-derived FVIII or recombinant FVIII (SIPPET, Survey of Inhibitors in Plasma-Product Exposed Toddlers). 41 Only rFVIII products produced in CHO and BHK cells were used (Recombinate Ò , Kogenate FS Ò , Advate Ò , and ReFacto AF Ò ).…”

Section: Recombinant Factor VIII Productsmentioning

confidence: 99%

Production of recombinant coagulation factors: Are humans the best host cells?

2017

View full text Add to dashboard Cite

The main treatment option for Hemophilia A/B patients involves the administration of recombinant coagulation factors on-demand or in a prophylactic approach. Despite the safety and efficacy of this replacement therapy, the development of antibodies against the coagulation factor infused, which neutralize the procoagulant activity, is a severe complication. The production of recombinant coagulation factors in human cell lines is an efficient approach to avoid such complication. Human cell lines can produce recombinant proteins with post translation modifications more similar to their natural counterpart, reducing potential immunogenic reactions. This review provides a brief overview of the most important characteristics of recombinant FVIII and FIX products available on the market and the improvements that have recently been achieved by the production using human cell lines.

show abstract

Characterisation of the post-translational modifications of a novel, human cell line-derived recombinant human factor VIII

Cited by 108 publications

References 60 publications

Life in the shadow of a dominant partner: the FVIII-VWF association and its clinical implications for hemophilia A

Life in the shadow of a dominant partner: the FVIII-VWF association and its clinical implications for hemophilia A

Alloantibodies to therapeutic factor VIII in hemophilia A: the role of von Willebrand factor in regulating factor VIII immunogenicity

Production of recombinant coagulation factors: Are humans the best host cells?

Contact Info

Product

Resources

About