Prediction of B- and T-cell epitopes using in-silico approaches: a solution to the development of recombinant vaccines against COVID-19

IntroductionMorus alba (white mulberry) pollen is an aero-allergen source that can trigger allergic diseases. Cobalamin-independent methionine synthase (MetE) in M. alba pollen has been proved to be one of the major allergens for some patients living in Istanbul (Turkey). The aim of the present study was the recombinant production and identification of MetE (Mor a 2), a novel allergen from M. alba pollen. The IgE binding reactivity of rMor a 2 produced for the first time was evaluated and some structural features were investigated by in silico methods to better understand its immunogenicity.Material and methodsThe gene encoding Mor a 2 was cloned in fission yeast, Schizosaccharomyces pombe ura4-D18h- strain, using pSLF1073 vector. This is the first report of the production of recombinant pollen allergen in S. pombe. After the purification, immunoreactivity of rMor a 2 was confirmed by immunoblotting using sera of patient allergic to M. alba pollen. Besides, B-cell epitopes of rMor a 2 were predicted using various bioinformatic tools, namely Bioinformatics Predicted Antigenic Peptides, BepiPred 2.0 and Immune Epitope Database whereas T-cell epitopes were estimated using NetMHCIIpan-3.2 and NetMHCII 2.3 servers.ResultsThe immunoblotting analysis yielded 11 of 11 positive reactions to rMor a 2. In silico predictions exerted seven B-cell epitopes (22-33, 384-394, 407-423, 547-553, 571-577, 671-678, 736-741) and seven T-cell epitopes (54-62, 161-170, 197-205, 347-358, 622-630, 657-665, 756-764).ConclusionsThese findings may help the use of rMor a 2 in the diagnosis and treatment of allergic diseases associated with M. alba and/or MetE.

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Rhipicephalus microplus lipocalins (LRMs): Genomic identification and analysis of the bovine immune response using in silico predicted B and T cell epitopes

International Journal for Parasitology ◽

10.1016/j.ijpara.2013.04.005 ◽

2013 ◽

Vol 43 (9) ◽

pp. 739-752 ◽

Cited By ~ 17

Author(s):

Manuel Rodriguez-Valle ◽

Paula Moolhuijzen ◽

Emily K. Piper ◽

Olivia Weiss ◽

Megan Vance ◽

...

Keyword(s):

Immune Response ◽

T Cell ◽

In Silico ◽

Rhipicephalus Microplus ◽

T Cell Epitopes

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Antibody Engineering of Anti-TFPI Bypass Therapeutic BAY 1093884: Isotype Selection and Sequence Optimization

Blood ◽

10.1182/blood.v126.23.3496.3496 ◽

2015 ◽

Vol 126 (23) ◽

pp. 3496-3496 ◽

Cited By ~ 5

Author(s):

Pedro Paz ◽

Jinger Xie ◽

Fred Aswad

Keyword(s):

T Cell ◽

Nk Cells ◽

Cell Lines ◽

In Silico ◽

Target Cells ◽

T Cell Epitopes ◽

Serum Complement ◽

Healthy Donors ◽

Bayer Healthcare

Abstract Background: BAY 1093884 is a fully human monoclonal antibody against tissue factor pathway inhibitor (TFPI) developed as a bypass agent for patients with hemophilia with or without inhibitors. It restores insufficient thrombin burst, leading to stable clot formation in hemophilic conditions in vitro, and effectively stops bleeding in vivo. Aims: Antibody targeting of TFPI provides the advantages of monoclonal antibodies, such as longer circulation half-life, and the possibility of subcutaneous administration. However, antibody therapeutics can present the challenge of delivering unwanted effects or activities, such as antibody-dependent cell cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) that target healthy tissues in the patient. Additionally, antibody sequence engineering that optimizes binding to the target protein may introduce immunogenic epitopes that can induce antidrug antibodies and compromise efficacy. Methods: In vitro assays used primary human umbilical vein endothelial cells and EA-hy926 cell lines as target cells. Total peripheral blood mononuclear cells (PBMCs) and purified natural killer (NK) cells isolated from healthy donors were used as effector cells for ADCC. Pooled human serum complement (Quidel®) was used for CDC assays. A proprietary immunogenicity scoring program for T-cell epitopes (Bayer HealthCare) was used to determine immunogenic potential of BAY 1093884 relative to antibodies in the market. Results: In vitro ADCC assessment of BAY 1093884 using PBMCs and NK cells isolated from healthy donors showed no lytic activity against TFPI-expressing primary and immortalized cell lines. Furthermore, no CDC activity using pooled serum complement was detected with BAY 1093884. In silico scoring for T-cell epitopes using a human leukocyte antigen DR-weighted algorithm compared favorably to therapeutic antibodies that have been reported to have low immunogenicity in the clinic. Conclusion: In vitro and in silico immunoprofiling has aided in the design of the bypass anti-TFPI immunoglobulin G2 antibody BAY 1093884 with low toxicity potential and immunogenicity in humans. Disclosures Paz: Bayer HealthCare: Employment. Xie:Bayer HealthCare: Employment. Aswad:Bayer HealthCare: Employment.

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