Patient anti-FVIII drug antibodies bind preferentially to a subset of FVIII covalent states.

Butera, D; Pijning, AE; Avery, NG; Coxon, CH; Metcalfe, C; Mimoun, A; Lacroix-Desmazes, S; Spiegel, PC; Hogg, PJ

Patient anti-FVIII drug antibodies bind preferentially to a subset of FVIII covalent states.

Butera, D Pijning, AE Avery, NG Coxon, CH Metcalfe, C Mimoun, A Lacroix-Desmazes, S Spiegel, PC Hogg, PJ

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Publisher:: ELSEVIER
Publication Type:: Journal Article
Citation:: Blood Adv, 2025, 9, (15), pp. 3706-3715
Issue Date:: 2025-08-12

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Field	Value	Language
dc.contributor.author	Butera, D
dc.contributor.author	Pijning, AE
dc.contributor.author	Avery, NG
dc.contributor.author	Coxon, CH
dc.contributor.author	Metcalfe, C
dc.contributor.author	Mimoun, A
dc.contributor.author	Lacroix-Desmazes, S
dc.contributor.author	Spiegel, PC
dc.contributor.author	Hogg, PJ
dc.date.accessioned	2026-01-22T05:04:48Z
dc.date.available	2025-03-20
dc.date.available	2026-01-22T05:04:48Z
dc.date.issued	2025-08-12
dc.identifier.citation	Blood Adv, 2025, 9, (15), pp. 3706-3715
dc.identifier.issn	2473-9529
dc.identifier.issn	2473-9537
dc.identifier.uri	http://hdl.handle.net/10453/192258
dc.description.abstract	Hemophilia A is a chronic life-threatening condition caused by the deficiency or dysfunction of plasma coagulation factor VIII (FVIII) and commonly managed by prophylaxis with regular infusion of FVIII protein. A major obstacle to FVIII replacement therapy is the generation of alloantibodies that diminish efficacy. Disulfide bonds link pairs of cysteine residues in proteins and, in several proteins, have been found to be only partially formed in the mature proteins. FVIII contains 8 disulfide bonds and their redox state in human blood and recombinant FVIII was determined using differential cysteine alkylation and mass spectrometry. All 8 disulfide bonds were found to be unformed in ∼10% to ∼70% of molecules of FVIII populations, which suggested a conformational flexibility that could favor the binding of certain ligands to subsets of FVIII with more or less formed disulfide bonds. To test this hypothesis, the binding of a panel of 5 patient-derived anti-FVIII antibodies to the population of FVIII disulfide-bonded states was evaluated. All 5 antibodies bound preferentially to FVIII states in which 2 or 3 of the 8 disulfides are significantly more unformed: C1918-C1922 in the A3 domain, C2040-C2188 in the C1 domain, and C2193-C2345 in the C2 domain. Disulfide bond mutagenesis experiments and molecular dynamics simulations indicate that this subset of FVIII states has long-range conformational dynamism that favors antidrug antibody binding. These findings will assist efforts to engineer an FVIII molecule that is less prone to neutralization by antidrug antibodies and has general implications for autoimmune conditions and antibody drug efficacy.
dc.format	Print
dc.language	eng
dc.publisher	ELSEVIER
dc.relation.ispartof	Blood Adv
dc.relation.isbasedon	10.1182/bloodadvances.2025016474
dc.rights	info:eu-repo/semantics/openAccess
dc.subject.classification	3201 Cardiovascular medicine and haematology
dc.subject.mesh	Humans
dc.subject.mesh	Factor VIII
dc.subject.mesh	Hemophilia A
dc.subject.mesh	Protein Binding
dc.subject.mesh	Disulfides
dc.subject.mesh	Protein Conformation
dc.subject.mesh	Humans
dc.subject.mesh	Hemophilia A
dc.subject.mesh	Disulfides
dc.subject.mesh	Factor VIII
dc.subject.mesh	Protein Conformation
dc.subject.mesh	Protein Binding
dc.subject.mesh	Humans
dc.subject.mesh	Factor VIII
dc.subject.mesh	Hemophilia A
dc.subject.mesh	Protein Binding
dc.subject.mesh	Disulfides
dc.subject.mesh	Protein Conformation
dc.title	Patient anti-FVIII drug antibodies bind preferentially to a subset of FVIII covalent states.
dc.type	Journal Article
utslib.citation.volume	9
utslib.location.activity	United States
pubs.organisational-group	University of Technology Sydney
pubs.organisational-group	University of Technology Sydney/Faculty of Science
pubs.organisational-group	University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	open_access	*
dc.rights.license	This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND 4.0). To view a copy of this license, visit https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.date.updated	2026-01-22T05:04:45Z
pubs.issue	15
pubs.publication-status	Published
pubs.volume	9
utslib.citation.issue	15

Abstract:

Hemophilia A is a chronic life-threatening condition caused by the deficiency or dysfunction of plasma coagulation factor VIII (FVIII) and commonly managed by prophylaxis with regular infusion of FVIII protein. A major obstacle to FVIII replacement therapy is the generation of alloantibodies that diminish efficacy. Disulfide bonds link pairs of cysteine residues in proteins and, in several proteins, have been found to be only partially formed in the mature proteins. FVIII contains 8 disulfide bonds and their redox state in human blood and recombinant FVIII was determined using differential cysteine alkylation and mass spectrometry. All 8 disulfide bonds were found to be unformed in ∼10% to ∼70% of molecules of FVIII populations, which suggested a conformational flexibility that could favor the binding of certain ligands to subsets of FVIII with more or less formed disulfide bonds. To test this hypothesis, the binding of a panel of 5 patient-derived anti-FVIII antibodies to the population of FVIII disulfide-bonded states was evaluated. All 5 antibodies bound preferentially to FVIII states in which 2 or 3 of the 8 disulfides are significantly more unformed: C1918-C1922 in the A3 domain, C2040-C2188 in the C1 domain, and C2193-C2345 in the C2 domain. Disulfide bond mutagenesis experiments and molecular dynamics simulations indicate that this subset of FVIII states has long-range conformational dynamism that favors antidrug antibody binding. These findings will assist efforts to engineer an FVIII molecule that is less prone to neutralization by antidrug antibodies and has general implications for autoimmune conditions and antibody drug efficacy.

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http://hdl.handle.net/10453/192258