Expressed truncated N-terminal variable surface glycoprotein (VSG) of Trypanosoma evansi in E. coli exhibits immuno-reactivity

Sengupta, P. P.; Balumahendiran, M.; Balamurugan, V.; Rudramurthy, Gudepalya Renukaiah; Prabhudas, K.

doi:10.1016/j.vetpar.2012.01.012

Cited by 26 publications

(18 citation statements)

References 24 publications

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“…evansi 11 . VSG has been expressed by several groups in different host cells including insect cell line 14 , yeast 15 and E. coli 16,17 . Furthermore, several other genes from different species of trypanosomes have also been successfully expressed in Pichia pastoris, for instance, acid -mannosidase and trans-sialidase from T. cruzi 18,19 , rhodesain from T. brucei rhodesiense 20 and congopain from T. congolense 21 .…”

mentioning

confidence: 99%

“…The isolates of T. evansi were propagated in rats 3 and purified 22 . The whole cell lysate (WCL) from different isolates were prepared 16 and the protein concentration of the supernatants was estimated 23 and kept in aliquots at -80C till further use.…”

mentioning

confidence: 99%

“…The hyper-immune/immune sera raised against different isolates 16 and available in the laboratory were used in the present study. However, the hyper immune serum against recombinant VSG (rVSG) was produced in New Zealand white adult rabbits following the protocol mentioned earlier 16 .…”

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confidence: 99%

“…However, the hyper immune serum against recombinant VSG (rVSG) was produced in New Zealand white adult rabbits following the protocol mentioned earlier 16 . The experimental animals were dealt following standard animal ethics and feed and drinking water were given ad libitum.…”

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confidence: 99%

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Development of ELISA Exploring Recombinant Variable Surface Glycoprotein for Diagnosis of Surra in Animals

et al. 2016

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Development of ELISA Exploring Recombinant Variable Surface Glycoprotein for Diagnosis of Surra in Animals

et al. 2016

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“…As a consequence, we cannot recommend Surra Sero K-SeT for testing of alpaca. It would be interesting to investigate whether the sub-optimal specificity of the Surra Sero K-SeT can be overcome by the use of alternative recombinant VSG RoTat 1.2 expressed in Escherichia coli or in Spodoptera frugiperda (Sengupta et al, 2012;Urakawa et al, 2001).…”

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confidence: 99%

Surra Sero K-SeT, a new immunochromatographic test for serodiagnosis of Trypanosoma evansi infection in domestic animals

Hadush

Rogé

Simon

et al. 2015

Veterinary Parasitology

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Trypanosoma evansi RoTat 1.2 variant surface antigen mimotopes selected by panning of the random peptide phage‐display library against monoclonal antibodies

Budania

Dubey

Singh

2022

J of Molecular Recognition

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Mimotope peptides of native antigens are valuable for diverse applications such as diagnostics, therapeutics and modern vaccine design. Here, we report for the first time the selection and identification of peptide mimotopes of Trypanosoma evansi RoTat 1.2 variant surface glycoprotein (VSG) for their potential uses in surra diagnostics and multi‐epitope vaccine research. First, we produced the mouse monoclonal antibodies (mAbs), designated as 2E11 (IgG1) and 1C2 (IgG1), against the antigens in T. evansi RoTat 1.2 lysates. We then used 2E11 mAb to immunoprecipitate the target antigen. The immunoprecipitated antigen was then identified to be the VSG by mass spectrometry. Both 2E11 and 1C2 mAbs reacted with the VSG in immunoblots. The surface plasmon resonance immunosensors developed with both the mAbs detected VSG in the parasite lysates as well as in the rodent sera. Further, the mAbs were biotinylated and used in three rounds of panning to select peptide mimotopes from the random peptide phage display library (PhD‐12; New England Biolabs, USA). The phage clones selected against each mAb were amplified and tested by phage capture ELISA for specificity. The peptide coding regions of the selected phages were sequenced and the protein blast search of the deduced amino acid sequences was performed by accessing the non‐redundant protein database at https://blast.ncbi.nlm.nih.gov/. The conformational B epitope prediction of the selected mimotope sequences was done by using 3D Pepitope algorithms accessed at: http://pepitope.tau.ac.il/. The potential applications of the selected mimotopes in surra diagnostics and research are being explored.

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Expressed truncated N-terminal variable surface glycoprotein (VSG) of Trypanosoma evansi in E. coli exhibits immuno-reactivity

Cited by 26 publications

References 24 publications

Development of ELISA Exploring Recombinant Variable Surface Glycoprotein for Diagnosis of Surra in Animals

Development of ELISA Exploring Recombinant Variable Surface Glycoprotein for Diagnosis of Surra in Animals

Surra Sero K-SeT, a new immunochromatographic test for serodiagnosis of Trypanosoma evansi infection in domestic animals

Trypanosoma evansi RoTat 1.2 variant surface antigen mimotopes selected by panning of the random peptide phage‐display library against monoclonal antibodies

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