An Update on Laboratory Diagnostics in Haemophilia A and B

Müller, Jens; Miesbach, Wolfgang; Prüller, Florian; Siegemund, T.; Scholz, Ute; Sachs, Ulrich J.

doi:10.1055/a-1665-6232

Cited by 17 publications

(50 citation statements)

References 111 publications

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“…Muller et al 13 result of these earlier experiences with rpFVIII and monitoring methods, it seems that in some laboratories the practice has been for a preference to use OSA despite both methods being available in some institutions. 15 One recent study has used porcine FVIII to prepare the calibration curve and compared it to HemosIL human plasma calibrator.…”

Section: Discussionmentioning

confidence: 99%

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Agreement between one stage and chromogenic assays in samples from patients receiving recombinant porcine FVIII (Obizur, Susoctocog‐alfa)

Guy,

Bowyer,

Shepherd

et al. 2023

Int J Lab Hematology

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IntroductionRecombinant porcine FVIII (rpFVIII) (Obizur, Susoctcog‐alfa, Takeda, Japan) is licensed for the treatment of bleeding in acquired Haemophilia A (AHA). The summary of product characteristics state that monitoring should be by one stage assay (OSA) rather than chromogenic assay (CSA). CSA have been shown to underestimate activity when rpFVIII is added to plasma in vitro.MethodsSamples from three AHA patients (n = 21) (pre‐ and post rpFVIII) were assessed using FVIII:C assays; OSA methods: Actin, Actin FS, Actin FSL and Pathromtin SL performed on CS5100i (Sysmex, Kobe, Japan); APTT‐SP, SynthASil and SynthAFax performed on ACL TOP (Werfen, Barcelona, Spain). CSA methods on CS5100i: Siemens Chromogenic Assay, Biophen FVIII:C, Technochrom FVIII:C; on ACL TOP: Rox Factor VIII, Coamatic Factor VIII and CRYOcheck Factor VIII.ResultsOSA and CSA varied according to reagent used median OSA 61 IU/dL (range 41.5–81 IU/dL (ANOVA p < 0.0001)) median CSA 46.5 IU/dL (range of method specific medians 36.5–84 IU/dL (ANOVA p < 0.0001)). Amongst OSA, Actin FS was associated with the highest FVIII:C, APTT‐SP was associated with the lowest. Variation in CSA results by different methods was also seen with highest FVIII:C levels obtained using the Technochrom FVIII:C and the lowest levels obtained with Siemens Assay.ConclusionThe relationship between OSA and CSA was not consistent between method or patient. Previously there has been reports of underestimation by CSA in in vitro spiked samples. Investigation into concentration of phospholipids in the APTT reagents may explain some of these variations.

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Section: Discussionmentioning

confidence: 99%

“…Muller et al 13 summarised the methods appropriate for testing rpFVIII based on previous publications, they break this down into individual APTT reagents and CSA methods. Actin FS, Actin FSL, APTT SP and SynthASil are deemed to be suitable, with insufficient data available for Actin and SynthAFax.…”

Section: Discussionmentioning

confidence: 99%

Agreement between one stage and chromogenic assays in samples from patients receiving recombinant porcine FVIII (Obizur, Susoctocog‐alfa)

Guy,

Bowyer,

Shepherd

et al. 2023

Int J Lab Hematology

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“…NBA‐derived inhibitor‐titers were calculated as described elsewhere 4 . However, instead of a fixed threshold of RA for positivity, run‐specific screening cut‐points (SCP) were determined as described below.…”

Section: Methodsmentioning

confidence: 99%

Two‐center validation of assays for the detection of binding and neutralizing anti‐factor VIII antibodies

Müller,

Neimanis,

Kahle

et al. 2023

Haemophilia

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IntroductionPatients with hemophilia A treated with coagulation Factor VIII (FVIII) products are at risk for developing anti‐FVIII antibodies. The ABIRISK Consortium aimed to provide knowledge on the formation and detection of anti‐drug antibodies against biopharmaceutical products, including FVIII. Accordingly, standardized and validated assays for the detection of binding (total) and neutralizing antibodies are needed.AimTwo‐center validation of an ELISA for the detection of total FVIII‐binding IgG‐antibodies and Nijmegen‐Bethesda assays for the quantification of FVIII‐neutralizing antibodies according to consensus validation guidelines.MethodsValidation of assays at both sites was done according to published recommendations and included preanalytics, the determination of key assay parameters, including cut‐points, assay sensitivity, precision, and FVIII interference.ResultsThe validated assays reproducibly detected FVIII‐binding and ‐neutralizing antibodies with comparable performance in both laboratories. Floating screening cut‐points were established for both assays. Determined mass‐based sensitivity of both assays (all values ≤66 ng/mL) complied with the minimum sensitivity for the detection of anti‐drug antibodies as recommended by the FDA (<100 ng/mL). Intra‐ and inter‐assay coefficients of variation did not exceed 25%. Assay validation further revealed that pre‐analytical heat treatment led to potentially false‐positive ELISA results, while up to 0.15 IU/mL, residual FVIII showed no significant impact. Overall, good agreement of results was found for patient samples analyzed at both study sites.ConclusionComprehensive validation of different anti‐FVIII‐antibody assays in two laboratories gave novel insights into the impact of pre‐analytical sample treatment as well as the comparability of test results generated by the use of methodically different assays.

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“…Moreover, as modifications in FVIII and FIX molecules in the extended half‐life (EHL) concentrates (fusion with the Fc fragment, albumin fusion and pegylation) alter the effect of FVIII and FIX in the laboratory tests, specific assays and reagents should be used for monitoring EHL‐products. Due to the significant differences between products, coagulation tests should be validated specifically for the particular EHL‐product 30 …”

Section: Update Of the European Guidelines For The Accreditation Of H...mentioning

confidence: 99%

“…Due to the significant differences between products, coagulation tests should be validated specifically for the particular EHL-product. 30 Furthermore, the non-factor product emicizumab, a bi-specific antibody mimicking the co-factor function of FVIIIa, artificially shortens Discrepancies in the assays measuring activities of FVIII and FIX following gene therapy have been also described, with OSCA results approximately 1.5 times higher than CSA in both haemophilia A and haemophilia B. 32,33 Therefore, laboratories providing testing for patients receiving gene therapy should be able to perform modified chromogenic test for the gene therapy program and follow-up of patients.…”

Section: Coagulation Laboratory Servicesmentioning

confidence: 99%

Accreditation model of European Haemophilia Centres in the era of novel treatments and gene therapy

Boban,

Baghaei,

Karin

et al. 2023

Haemophilia

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IntroductionThe international certification of haemophilia centres in Europe is run by the European Association of Haemophilia and Allied Disorders (EAHAD) and European Haemophilia Consortium (EHC) since 2013. The centres are designated as European Haemophilia Comprehensive Care Centres (EHCCC) or European Haemophilia Treatment Centres (EHTC), based on the specific requirements which evaluate centres’ ability to provide care for patients with haemophilia and allied disorders.AimTo establish the new protocol for accreditation of European Haemophilia Centres.MethodsEAHAD, in collaboration with EHC, established Accreditation Working Group with the aim to define necessary measures to safeguard quality and improvement of bleeding disorders care throughout Europe and to build a novel model for accreditation of European Haemophilia Centres.ResultsThe European guidelines for certification of haemophilia centres have been updated to guidelines for the accreditation and include all the requirements regarding facilities, laboratory and personnel needed for optimal management of novel treatment options, including the introduction of the hub‐and‐spoke model for delivery of gene therapy. A pilot project for the accreditation of haemophilia centres including on‐site audit has been designed.ConclusionImplementation of the novel accreditation protocol of the haemophilia treatment and haemophilia gene therapy centres has been made to further improve the quality of care for patients with haemophilia and other inherited bleeding disorders.

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An Update on Laboratory Diagnostics in Haemophilia A and B

Cited by 17 publications

References 111 publications

Agreement between one stage and chromogenic assays in samples from patients receiving recombinant porcine FVIII (Obizur, Susoctocog‐alfa)

Agreement between one stage and chromogenic assays in samples from patients receiving recombinant porcine FVIII (Obizur, Susoctocog‐alfa)

Two‐center validation of assays for the detection of binding and neutralizing anti‐factor VIII antibodies

Accreditation model of European Haemophilia Centres in the era of novel treatments and gene therapy

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