scholarly journals Factor VIII Epitope Analysis Using a Random Peptide Phage-Display Library Approach in the Sippet Cohort

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 3176-3176
Author(s):  
Shermarke Hassan ◽  
Guido Baselli ◽  
Kaia Palm ◽  
Frits Richard Rosendaal ◽  
Roberta Palla ◽  
...  
Keyword(s):  
Phage Display ◽  
Factor Viii ◽  
Epitope Mapping ◽  
Hemophilia A ◽  
Phage Display Library ◽  
Mapping Technique ◽  
Discriminative Performance ◽  
Inhibitor Development ◽  
Random Peptide ◽  
Peptide Phage Display

Abstract Background Inhibitor development is the most severe complication of hemophilia A care, and is associated with increased morbidity and mortality. Aims The aim of this study was to use a novel epitope mapping method to explore the factor VIII (FVIII)-specific epitope profile in the SIPPET cohort population. Methods The population consisted of 122 previously untreated patients with severe hemophilia A that were followed-up for 50 days of exposure to FVIII. Sampling was performed before FVIII treatment and at the end of the follow-up. The outcome was inhibitor development. The FVIII-specific IgG epitope repertoire was assessed by means of a novel high-throughput epitope mapping technique using a random peptide phage-display library. Using this assay, a set of affinity-selected 12-mer peptide sequences (also called mimotopes) that were strongly bound by FVIII-specific antibodies were identified. These mimotopes were clustered on the basis of sequence similarity and a consensus motif was generated for each mimotope cluster. Discriminative performance of these mimotope clusters was assessed by ROC analysis. Mimotope clusters were mapped onto the 3D structure of a B-domain deleted FVIII model using a B-cell epitope prediction algorithm (Mapitope). Results The FVIII-specific antibody response is polyclonal with several mimotope clusters. The most predominant mimotope clusters in inhibitor patients were mapped to the heavy chain of the FVIII molecule. Using plasma samples taken before exposure to FVIII, three mimotopes (with the consensus motifs "QM", "PSLxWK" and "SWPHxxxxK") were identified that predicted inhibitor development (with an AUC of 0.76, 0.80 and 0.76 respectively). Conclusion Information on immunodominant epitope clusters can be used to generate novel, less immunogenic FVIII proteins and set up diagnostic tests that predict the risk of inhibitor development before starting treatment with FVIII. Figure 1 Figure 1. Disclosures Palm: Protobios LLC: Current Employment, Patents & Royalties: Inventor of the patent application (PCT Application No. US/14079626) filed by Protobios that covers the use of phage display method to manipulate and monitor humoral immunity.. Palla: Pfizer: Other: Travel support; Kedrion: Other: Travel support; Novonordisk: Speakers Bureau. Peyvandi: Roche: Consultancy, Honoraria; Sanofi: Consultancy, Honoraria; Sobi: Consultancy, Honoraria; Takeda: Consultancy, Honoraria.

scholarly journals Epitope mapping by random peptide phage display reveals essential residues for vaccinia extracellular enveloped virion spread

2012 ◽  
Vol 9 (1) ◽  
pp. 217 ◽  
Author(s):  
Yong He ◽  
Yonggang Wang ◽  
Evi B Struble ◽  
Pei Zhang ◽  
Soma Chowdhury ◽  
...  
Keyword(s):  
Phage Display ◽  
Epitope Mapping ◽  
Random Peptide ◽  
Peptide Phage Display

Epitope mapping of polyclonal clotting factor VIII-inhibitory antibodies using phage display

2004 ◽  
Vol 91 (03) ◽  
pp. 619-625 ◽  
Author(s):  
Christiane Mühle ◽  
Stefan Schulz-Drost ◽  
Alexey Khrenov ◽  
Evgueni Saenko ◽  
Jens Klinge ◽  
...  
Keyword(s):  
Phage Display ◽  
Factor Viii ◽  
Inhibitory Activity ◽  
Phage Display Library ◽  
Clotting Factor ◽  
C2 Domain ◽  
Clotting Factors ◽  
Antibody Preparation ◽  
Random Peptide ◽  
Inhibitory Antibodies

SummaryClotting factor VIII (fVIII)-inhibitory antibodies represent a major problem in the treatment of haemophilia A. To understand the inactivation mechanisms and to pave the way towards modifications of recombinant clotting factors that reduce their immunogenicity, the exact localization of immunodominant epitopes is required. Here, a random peptide phage display library was employed to identify epitopes of polyclonal fVIII antibodies isolated from patient’s plasma by affinity chromatography. FVIIIbinding specificity and inhibitory activity of the isolated fVIII antibodies were confirmed by ELISA and Bethesda assays. Phage selection on the individual samples yielded several phages which were displaced from binding to the respective antibody preparation by fVIII. Their homology with amino acid motifs of human fVIII and immunoprecipitation results with radioactively labelled fVIII fragments suggested putative epitopes in the A1, A2 and C1 domains of fVIII for one and in the C2 domain for another patient. Synthetic peptides corresponding to the A2, C1 and C2 domain epitopes blocked antibody binding to fVIII and partially neutralized the inhibitory activity of the respective plasma in Bethesda assays. These results provide the proof of principle that random peptide libraries can be used for the mapping of epitopes in a polyclonal antibody preparation.

scholarly journals Boophilus microplus cathepsin L-like (BmCL1) cysteine protease: Specificity study using a peptide phage display library

2011 ◽  
Vol 181 (2-4) ◽  
pp. 291-300 ◽  
Author(s):  
Renan O. Clara ◽  
Tatiane S. Soares ◽  
Ricardo J.S. Torquato ◽  
Cássia A. Lima ◽  
Renata O.M. Watanabe ◽  
...  
Keyword(s):  
Phage Display ◽  
Cysteine Protease ◽  
Cathepsin L ◽  
Phage Display Library ◽  
Boophilus Microplus ◽  
Peptide Phage Display ◽  
Specificity Study ◽  
Protease Specificity

scholarly journals Molecular and clinical predictors of inhibitor risk and its prevention and treatment in mild hemophilia A

Blood ◽  
2014 ◽  
Vol 124 (15) ◽  
pp. 2333-2336 ◽  
Author(s):  
Giancarlo Castaman ◽  
Karin Fijnvandraat
Keyword(s):  
Factor Viii ◽  
Hemophilia A ◽  
Biological Response ◽  
Interindividual Variation ◽  
Clinical Use ◽  
Clinical Predictors ◽  
Inhibitor Development ◽  
Prevention And Treatment ◽  
Selection Of

Abstract The risk for inhibitor development in mild hemophilia A (factor VIII levels between 5 and 40 U/dL) is larger than previously anticipated, continues throughout life, and is particularly associated with certain mutations in F8. Desmopressin may reduce inhibitor risk by avoiding exposure to FVIII concentrates, but the heterogenous biological response to desmopressin, showing large interindividual variation, may limit its clinical use. However, predictors of desmopressin response have been recently identified, allowing the selection of the best candidates to this treatment.

scholarly journals Identification of Efficiently Cleaved Substrates for HIV-1 Protease Using a Phage Display Library and Use in Inhibitor Development

Virology ◽  
2000 ◽  
Vol 274 (2) ◽  
pp. 391-401 ◽  
Author(s):  
Zachary Q. Beck ◽  
Laurence Hervio ◽  
Philip E. Dawson ◽  
John H. Elder ◽  
Edwin L. Madison
Keyword(s):  
Phage Display ◽  
Phage Display Library ◽  
Inhibitor Development ◽  
Hiv 1

THE RISK OF INHIBITOR DEVELOPMENT IN 1104 PATIENTS WITH HEMOPHILIA A ACCORDING TO FACTOR VIII GENOTYPE

2007 ◽  
Vol 5 ◽  
pp. O-S-064-O-S-064
Author(s):  
J. Boekhorst ◽  
G. Rastegar Lari ◽  
R. d'Oiron ◽  
J.M. Costa ◽  
I.R.O. Novakova ◽  
...  
Keyword(s):  
Factor Viii ◽  
Hemophilia A ◽  
Inhibitor Development

scholarly journals Incidence of inhibitor development in a group of young hemophilia A patients treated exclusively with lyophilized cryoprecipitate

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3332-3335 ◽  
Author(s):  
K Peerlinck ◽  
FR Rosendaal ◽  
J Vermylen
Keyword(s):  
Factor Viii ◽  
Patient Group ◽  
Hemophilia A ◽  
Cumulative Risk ◽  
Reference Population ◽  
Single Factor ◽  
Inhibitor Development ◽  
Age Dependent ◽  
The One ◽  
Suitable Reference

Abstract The incidence of neutralizing isoantibody formation to infused factor VIII in a cohort of 67 hemophilia A patients, born between January 1, 1971 and April 30, 1990, who had been treated exclusively with lyophilized cryoprecipitate, was 6% (5.3 per 1,000 patient years of observation). The age-dependent cumulative risk was 4.6% at 4 years of age and 6.7% at 8 years of age. Recent reports in patients treated with a variety of more pure concentrates show a much higher incidence of inhibitor formation and tend to be used as a reference when new concentrates are introduced. We believe that a patient group, such as the one studied here, is a more suitable reference population because these patients have been exclusively treated with a single factor VIII preparation.

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