Figure 6

Enhanced immune response after immunisation with gp41 mutated in the isu domain. (A) Comparative ELISA of sera from two rats immunised with the wt gp41 and with the mutated gp41(Q2A). T20 (100 ng per well) was used as antigen. (B) Comparative ELISA of sera from rats immunised with the mutated gp41ΔCT(Q2A) (rats 1–3) and rats immunised with the wt gp41ΔCt (rats 4–6), all without the cytoplasmatic tail and the membrane spanning domain. 100 ng of T20 per well were used as antigen. Sera were diluted 1–1:200. 2–1:400, 3–1:800, 4–1:1600. (C) Western blot analysis of the rat sera using the recombinant ectodomain of gp41 (17 kDa) produced in bacteria. Lane 1, 2 - pre immune sera from rats 1 and 6, lane 3 - mab 2F5 (positive control), lanes 4–6 - sera from rats 1, 2 and 3, all immunized with mutated gp41ΔCt (Q2A), lanes 7–8 - sera from rats 5 and 6, both immunized with wt rgp41ΔCT. The rat sera were diluted 1:200, the mab 2F5 was used at 2 μg/ml, rabbit anti-rat and rabbit anti-human sera conjugated with alkaline phosphatase at 1:2000.