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Figure 2: Analysis of the LTB-HIV gag p24 conjugate.
Conjugation of HIV gag p24 to LTB was performed by using the bifunctional crosslinker, SPDP. (A) The conjugate was dialyzed against PBS pH 7.4, loaded and run on 10% protein gel under non-reducing condition. The gel shows 2 bands, a lower band of approximately 100 KD and a higher band on the top of the gel of approximately 200 KD. (B) A GM1-ELISA was performed for the analysis of the conjugate. The conjugate was probed on GM1-coated wells. The conjugate was serially diluted (2x) and probed with mouse anti-gag p24 and goat anti-mouse alkaline phosphatase. The positive control consisted of mouse anti-gag p24 and goat anti-mouse alkaline phosphatase incubated in wells coated with gag p24 (p24 control). The negative control consisted of GM1 wells incubated with LTB-gag p24 conjugate and goat anti-mouse alkaline phosphatase in the absence of the primary antibody. The reaction was developed by addition of alkaline phosphatase substrate and read at 405 nm. |