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Recombinant Proteins> | … 1B, white microtitre plates (Corning, USA) were coated with 5 μg/ml of protein G (Genscript Co., China) in 0.01 M PBS and incubated overnight at 4 °C. The plates were then washed five times with 0.01 M PBS containing 0.05% Tween 20 (PBS-T) and blocked with 5% non-fat … | Get A Quote |
In this study, we described an ultrasensitive and high-throughput luciferase immunosorbent assay (LISA) for qualitative and quantitative detection of anti-HIV-1 antibody. Anti-HIV antibody in serum or plasma samples was captured by protein A/G-coated microtiter plate and detected with crude cell lysates expressing Nanoluc luciferase (Nluc) enzyme fused with HIV-1 p24 or gp41 antigen without the need of protein purification. After the addition of furimazine substrate, anti-HIV antibodies were quantitatively measured as luciferase light units. LISA showed a wide linear range of detection and was about 10-fold more sensitive than ELISA. For the detection of both anti-p24 and anti-gp41, LISA showed extraordin... More