PXD060297-1

PXD060297 is an original dataset announced via ProteomeXchange.

Title	Patient anti-FVIII drug antibodies bind preferentially to a subset of FVIII covalent states
Description	Haemophilia A is a chronic life-threatening condition caused by deficiency or dysfunction of plasma coagulation factor VIII (FVIII) and prophylaxis by regular infusion of FVIII protein is a common treatment. 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 disulphide bonds were found to be unformed in 1 in 10 to 2 in 3 molecules of FVIII populations, which suggested a conformational flexibility that could favour 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 five patient-derived anti-FVIII antibodies to the population of FVIII disulfide-bonded states was evaluated. All five antibodies bound preferentially to FVIII states where 2 or 3 of the 8 disulphides 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 have long-range conformational dynamism that favours anti-drug antibody binding. These findings will assist efforts to engineer a FVIII molecule that diminishes the emergence of inhibitors and has general implications for autoimmune conditions and antibody drug efficacy.
HostingRepository	PRIDE
AnnounceDate	2025-08-04
AnnouncementXML	Submission_2025-08-03_16:11:50.130.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Diego Butera
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2025-01-28 15:09:00	ID requested
⏵ 1	2025-08-03 16:11:50	announced

10.1182/bloodadvances.2025016474;

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. Blood Adv, 9(15):3706-3715(2025) [pubmed]

submitter keyword: Plasma, mass spec, FVIII

Philip John Hogg
contact affiliation	School of Life Sciences, University of Technology Sydney and Centenary Institute, University of Sydney, Sydney NSW, Australia
contact email	phil.hogg@sydney.edu.au
lab head
Diego Butera
contact affiliation	ACRF Centenary Cancer Research Centre
contact email	diegobutera@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD060297
     1. Label: PRIDE project
     2. Name: Patient anti-FVIII drug antibodies bind preferentially to a subset of FVIII covalent states