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. 2021 Jul 1;11:708739. doi: 10.3389/fcimb.2021.708739

Figure 2.

Figure 2

Analysis of rLIC13086 by SDS-PAGE, CD Spectroscopy, and serum reactivity with serum of confirmed leptospirosis. (A) Recombinant protein purification samples were separated by SDS-PAGE (12%) and stained with Comassie Blue. M: molecular weight LMW (Ge Healthcare), (1) non-induced total lysates, (2) induced total lysates, (3) induced soluble fraction, (4) induced insoluble fraction, (5–8) washing steps with imidazole (5, 20, 40, and 60 mM), and (9) purified protein after elution. (B) CD spectra was recorded as accumulative of 10 scans from 260–190 nm at 25°C and data are expressed in molar residual ellipticity. (C) The recombinant protein was immobilized onto ELISA plates (250 ng/well), and the reactivity was assessed by total IgG antibodies in sera from leptospirosis patients at convalescent phase (MAT+) and detection with incubation of HRP-conjugated anti-human IgG (1:5,000). The cutoff values are represented by the horizontal line and defined as the mean plus three standard deviation of the absorbances values from normal human serum (NHS).