Dataset of selection and identification of mimotopes of the variant surface glycoprotein of Trypanosoma evansi RoTat1.2 isolate by PhD-12 peptide phage display library panning against monoclonal antibodies

Published: 7 April 2021| Version 2 | DOI: 10.17632/bs6pbskc8n.2
Contributors:
,
Savita Budania

Description

By employing two murine monoclonal antibodies (mAbs), designated as 2E11 (IgG1) and 1C2 (igG1), we have selected and identified the peptide mimotopes of T. evansi RoTat 1.2 variant surface glycoprotein (VSG) [File 1]. The mAbs-selected mimotopes were then used to predict the conformational B epitopes on VSG. First, we produced by the hybridoma technique the mAbs against the T. evansi RoTat 1.2 lysate antigens. The 2E11 mAb was used to immunoprecipitate the target antigen in the parasite lysate, which was then identified to be the VSG by mass spectrometry. Both 2E11 and 1C2 mAbs reacted with the VSG in the Western blots. Further, the biotinylated mAbs were used in three rounds of panning to select the peptide mimotopes out of the phage-displayed 12-mer random peptides library (PhD-12; New England Biolabs, USA) [File 1-4; Fig. 1-2]. The peptide-displaying phage clones specifically bound to the selector mAb in the phage- capture ELISA [File 4 table 6-7]. The peptide coding regions of the selected phages were sequenced [File 4 table 4-5] and the related sequences were blast searched in the non-redundant protein database at https://blast.ncbi.nlm.nih.gov/ [File 5-13]. By using 3D Pepitope algorithms accessed at http://pepitope.tau.ac.il/, the mimotope sequences predicted the conformational B epitopes on T. brucei IlTat1.24 VSG (PDB id: 2VSG_A or 2VSG_B) [File 14-28; Fig. 3-4]. T. brucei IlTat1.24 2VSG_A and 2VSG_B showed homology to T. evansi RoTat1.2 VSGs of various isolates [File 29-35]. Data generated from our laboratory experiments and by application of bioinformatics tools are presented for the B epitope mapping of the VSG of an Indian isolate of Trypanosoma evansi RoTat1.2. Four different peptide mimotope sequences (deduced amino acid sequences) were obtained from 11 sequences of 2E11-binder clones (File 4 table 4]. The consensus mimotope [HWKAVNWLKPWT] was represented by 8 identical sequences. Whereas, five distinct mimotope sequences were obtained from 14 1C2-binder clones. While the consensus sequence [WHWDANRWWTYR] was represented by 9 identical sequences [File 4 table 5]. The conformational B epitopes were predicted by Pepsurf alignment of 2E11 and 1C2 mAb- selected peptide mimotopes against T. brucei IlTat1.24 VSG (PDB ID: 2VSG_A) using 3D Pepitope algorithms [accessed at: (http://pepitope.tau.ac.il/)] [File 14-28; Fig. 3-4]. The best cluster identified by three 2E11-selected mimotopes consisted of 14 AA [File 24], whereas that identified by two 1C2-selected mimotopes consisted of 17 AA [File 26]. Of these AA in the best clusters, nine AA, i.e., T40, P259, T262, V264, T266, E314, A315, W317 and N318 were found to be shared or common. The conformational B cell epitopes on 2VSG_A predicted by the DiscoTope tool at http://www.iedb.org/ shared some AA positions in both 2E11 and 1C2-selected mimotopes [Fig. 4 & File 36]. These data could be useful for diagnostics, vaccine design, and research in trypanosome biology.

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By employing two murine monoclonal antibodies (mAbs), designated as 2E11 (IgG1) and 1C2 (igG1), we have selected and identified the peptide mimotopes of T. evansi RoTat 1.2 variant surface glycoprotein (VSG). The mAbs-selected mimotopes were then used to predict the conformational B epitopes on VSG. To achieve that, we first produced by the hybridoma technique the mAbs against the T. evansi RoTat 1.2 lysate antigens. The 2E11 mAb was used to immunoprecipitate the target antigen in the parasite lysate, which was then identified to be the VSG by mass spectrometry. Both 2E11 and 1C2 mAbs reacted with the VSG in the Western blots. Further, the biotinylated mAbs were used in three rounds of panning to select the peptide mimotopes out of the phage-displayed 12-mer random peptides library (PhD-12; New England Biolabs, USA). The peptide-displaying phage clones specifically bound to the selector mAb in the phage- capture ELISA. The peptide coding regions of the selected phages were sequenced and the related sequences were blast searched in the non-redundant protein database at https://blast.ncbi.nlm.nih.gov/. By using 3D Pepitope algorithms accessed at http://pepitope.tau.ac.il/, the mimotope sequences predicted the conformational B epitopes on T. brucei IlTat1.24 VSG (PDB id: 2VSG_A or 2VSG_B). T. brucei IlTat1.24 2VSG_A and 2VSG_B showed homology to T. evansi RoTat1.2 VSGs of various isolates. Data generated from our laboratory experiments and by application of bioinformatics tools are presented for the B epitope mapping of the VSG of an Indian isolate of Trypanosoma evansi RoTat1.2.