Activity of transgene-produced B-domain-deleted factor VIII in human plasma following AAV5 gene therapy.

Adeno-associated virus (AAV)-based gene therapies can restore endogenous factor VIII (FVIII) expression in hemophilia A (HA). AAV vectors typically use a B-domain-deleted FVIII transgene, such as human FVIII-SQ in valoctocogene roxaparvovec (AAV5-FVIII-SQ). Surprisingly, the activity of transgene-produced FVIII-SQ was between 1.

FIX potency of rFIX-Albumin fusion protein is underestimated by one-stage methods using silica-based APTT reagents.

Introduction: Higher potency is obtained with chromogenic substrate (CS) methods and one-stage (OS) method with SynthAFax vs silica-based OS methods on analysis of albutrepenonacog alpha (rFIX fused with albumin, rFIX-FP).

Aim: Investigation of the effect of contact activator in search for explanation of discrepancy between methods.

Methods: Chromogenic Rox Factor IX method and OS methods with Pathromtin SL, SynthAFax or new OS method variants using different phospholipid emulsions and addition of either colloidal silica to create APTT reagents or addition of human FXIa together with calcium ions, in the latter case omitting contact activation.

New Tools to Study Contact Activation.

The recent availability of a sensitive chromogenic method approach for determination of FXIa activity has been explored for designing sensitive methods for FXIIa and kallikrein, both using FXa formation as the read-out. For both enzymes the assay range 1-10 nmol/L provides a resolution of about 0.8 absorbance units with a total assay time of about 20 min.

Characterization of thrombin derived from human recombinant prothrombin.

Thrombin (FIIa) is the key enzyme in haemostasis and acts on several substrates involved in clot formation, platelet activation and feed-back regulation of its own formation. During activation of blood coagulation, FIIa is formed by proteolytic cleavage of prothrombin (FII). In the production of recombinant human FII (rhFII), a key question is whether the thrombin formed has the same properties as endogenous thrombin.

Criteria for specific measurement of plasminogen (enzymatic; procedure) in human plasma.

There is a lack of well-established criteria for the specific measurement of fibrinolytic variables. On behalf of the SSC the Subcommittee on Fibrinolysis started a process to develop such criteria. This report describes the criteria for the measurement of plasminogen (enzymatic; procedure) in human plasma.