Functional Insights into Recombinant TROSPA Protein from Ixodes ricinus
Figure 2
Comparison of the relative level of OspA protein from different Borrelia species bound to TROSPA.
TROSPA protein-coated ELISA microplates were incubated with OspA protein (c = 30 µg/ml) from B. afzelii, B. garinii or B. burgdorferi s. s. Bound OspA was subsequently detected using rabbit Borrelia-specific polyclonal IgG (primary antibody) and AP-conjugated anti-rabbit polyclonal IgG (secondary antibody). Absorbance of the soluble product of the alkaline phosphatase reaction was measured with reference to blank samples containing no OspA. As a control, analogous assays with OspC from B. garinii were performed. The level of individual OspA protein binding was always determined based on 16 reactions (for details see Materials and methods). For each protein pair arithmetic mean of absorbance and SD was calculated and expressed in percentage, assuming that the level of TROSPA binding with OspA from B. garinii is 100%. The differences between bound OspA from B. garinii and OspA from B. afzelii (p = 0,0003) or OspA from B. garinii and OspA from B. burgdorferi s. s. (p<0,0001) were statistically significant.