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Kinetics of the interaction between anti‐FVIII antibodies and FVIII from therapeutic concentrates, with and without von Willebrand factor, assessed by surface plasmon resonance
Author(s) -
Grancha S.,
Ortiz A. M.,
Marañón C.,
Hampel K.,
Moret A.,
Zimmermann B.,
Jorquera J. I.,
Aznar J. A.
Publication year - 2012
Publication title -
haemophilia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.213
H-Index - 92
eISSN - 1365-2516
pISSN - 1351-8216
DOI - 10.1111/j.1365-2516.2012.02858.x
Subject(s) - von willebrand factor , surface plasmon resonance , immunogenicity , haemophilia a , haemophilia , recombinant dna , antibody , chemistry , kinetics , dissociation (chemistry) , biophysics , medicine , immunology , biochemistry , platelet , surgery , materials science , nanotechnology , biology , physics , quantum mechanics , nanoparticle , gene
Summary The presence of VWF in plasma‐derived FVIII (pdFVIII/VWF) products has been pointed out as a key difference with recombinant FVIII ( rFVIII ) products with regard to immunogenicity. A Surface Plasmon Resonance (SPR) study was designed to characterize in detail the interaction between anti‐FVIII (IgGs) from a severe haemophilia A patient, and FVIII from concentrates of different sources. Full‐length rFVIII (preincubated or not with purified VWF), B domain‐deleted (BDD)‐ rFVIII and pdFVIII/VWF were analysed. To ensure reproducible conditions for accurate determination of kinetic constants, a capture‐based assay format was developed using protein G surfaces for specific and reversible coupling of endogenous anti‐FVIII antibodies. Concentration ranges (n m ) of FVIII products tested were 9–0.03 ( rFVIII ) and 6–0.024 (pdFVIII/VWF). The association with antibodies was monitored for 3–5 min, whereas dissociation of the complex was followed for 5–20–240 min. A strong interaction of rFVIII and BDD‐ rFVIII with patient's IgG was detected with the K D values in the low picomolar range (5.9 ± 3.0 and 12.7 ± 6.9 p m , respectively) and very slow dissociation rates, while pdFVIII/VWF showed only marginal binding signals. The VWF complexed rFVIII displayed reduced binding signals compared with uncomplexed rFVIII , but the K D was still in the picomolar range (4.1 ± 1.9 p m ) indicating insufficient complex formation. rFVIII , alone or bound to exogenously added VWF, showed high affinity for anti‐FVIII IgGs from a severe haemophilia A patient whereas pdFVIII/VWF did not. These results are in agreement with those studies that point towards rFVIII concentrates to be more immunogenic than pdFVIII concentrates.