Surface plasmon resonance dataset of Trypanosoma evansi RoTat1.2 variant surface glycoprotein antigen interaction with monoclonal antibody- coated biosensors
Description
The surface plasmon resonance (SPR) bio-(immuno-)sensors are being developed for the diagnosis of infectious diseases, cancers, food safety, etc. The SPR immunosensor using a monoclonal antibody as the capture biomolecule coated onto the gold chip allows direct, rapid, real-time, label-free, quantitative and cost-effective detection of the target antigens as analyte in a test sample. We developed for the first time SPR immunosensors using two monoclonal antibodies, viz., 2E11 (IgG1) and 1C2 (IgG1), produced in our laboratory for real-time, label-free, and rapid detection of their target antigen, i.e., Trypanosoma evansi RoTat 1.2 variant surface glycoprotein (VSG) in sera samples from the laboratory rodents and the field bovines [File 1 pdf]. First, we produced by the hybridoma technique several mAbs that reacted with the T. evansi RoTat 1.2 lysate Ags [File 2 pdf]. One of these mAbs, viz., 2E11 mAb was then used to immunoprecipitate the target Ag in the parasite lysate [Fig. 1A & 1B; File 3 pdf], which was then identified as T. evansi VSG by mass spectrometry [Fig. 2; File 3 pdf & File 4 Excel data]. Both 2E11 and 1C2 mAbs reacted with the VSG Ag in the Western blots [Fig. 3; File 3 pdf]. Then, the interactions of these mAbs with the above VSG Ag in the parasite lysate were analyzed by the respective SPR-immunosensor. The immunosensor was developed by binding of the biotinylated mAbs onto streptavidin immobilized on the gold chip [Dutra, RF and Kubota, LT. (2006). Clinica Chimica Acta. 379, 114-120]. The equilibrium dissociation constants (KD= kd/ka) of mAbs-VSG were determined to be 127 nM (ka=196.4 ± 61.9 s-M-; kd=2.51E-05 s-) for 2E11 mAb and 290 pM (ka=4616.1 ± 170.1 s-M-; kd=1.36E-06 s-) for 1C2 mAb (Table 1 pdf; Files 5 & 6 Excel data; Fig. 4-5 pdf). Further, we produced the SPR data and the sensograms of the interactions of 2E11 and 1C2 mAbs with the VSG Ag in the sera samples of the parasite-infected laboratory rodents as well as the test sera samples from the field cattle and buffaloes [File 7 with Fig. 6-11 Excel data; Fig. 6-11 pdf; File 8 with Fig. 13-17 Excel data; Fig. 12-17 pdf]. In addition, the kinetic parameters of the mAb interactions with two synthetic peptide mimotopes of the VSG were determined (Table 1 pdf; File 9 with Fig. 18-21 Excel data; Fig. 18-21 pdf). The peptide mimotopes of VSG were previously selected by PhD-12 phage display library panning against 2E11 and 1C2 mAbs in another study [Dataset reference: Mendeley Data, V2, doi: 10.17632/bs6pbskc8n.2]. These data provide valuable information for developing the real-time, label-free, SPR- based immunosensors for the diagnosis of surra caused by Trypanosoma evansi infection in a wide variety of domestic, zoo, and wildlife animal species.
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We developed for the first time SPR immunosensors using two monoclonal antibodies, viz., 2E11 (IgG1) and 1C2 (IgG1), produced in our laboratory for real-time, label-free and rapid detection of their target antigen, i.e., Trypanosoma evansi RoTat 1.2 variant surface glycoprotein (VSG) in sera samples from the laboratory rodents and the field bovines. First, we produced by the hybridoma technique several mAbs that reacted with the T. evansi RoTat 1.2 lysate Ags. Then, 2E11 mAb was used to immunoprecipitate the target Ag in the parasite lysate. The 2E11-precipitated Ag was identified as T. evansi VSG by Mass spectrometry. Both 2E11 and 1C2 mAbs reaction with the VSG Ag was detected by Western blotting. The real-time label-free interactions between each mAb and the parasite Ag in the buffer or serum samples were analyzed by the SPR method using a dual-channel kinetic evaluation instrument (KEI) (KE Instruments BV ESPIRIT®, The Netherlands). All the experiments were performed at 25°C. The interactions between the gold surface- streptavidin-bound mAbs (2E11 and 1C2, separately) and the parasite antigen in the test samples were recorded as a differential of SPR responses in channel 1 (the test) and channel 2 (the reference). The immunosensor response data acquisition software version 4.4 and the Kinetic evaluation software version 5.4 were provided with the system. To develop the immunosensors, the mAbs were biotinylated and made to bind to streptavidin-immobilized onto the gold chip according to Dutra & Kubota(2006) (Clinica Chimica Acta. 379, 114-120). Briefly, each biotinylated mAb (10µg/ml in PBS, pH7.4), was captured on streptavidin onto the gold disk, and the non-reacting sites were blocked by 10mM glycine in PBS. The KD values of the mAbs were determined using a range of concentrations of the purified T. evansi lysate antigen. After each run, the immunosensor surface was regenerated with 0.1M NaOH in PBS. Kinetic parameters of the Ag-mAb interactions, i.e., association and dissociation rate constants, ka and kd, respectively, were calculated from the association phase of the sensograms using the software supplied with the instrument. The equilibrium dissociation constant, KD, was calculated from the ratio of kd/ka. Next, a total of 50 field bovine sera samples were analyzed by each SPR immunosensor system for detection of the parasite Ag, taking fetal bovine serum (FBS) as the negative control. Each serum sample was diluted 1:10 in PBS and 50 µl run over each immunosensor for obtaining the SPR sensogram. A proportionate shift in SPR response was taken as the indicator of the Ag-mAb interaction. The SPR dataset and the sensograms produced by the mAbs with the rodent and the bovine sera samples were compiled and presented [File 7 & 8]. In addition, SPR data was also acquired to analyse the interactions of 2E11 and 1C2 immunosensors with in-house synthesized peptide VSG mimotopes, HWKAVNWLKPWK (M-12), and HWDANRAWTYR (M-11) [Table 1; File 9].