Alkaline phosphatase conjugated anti human igg antibody

Polystyrene plates were coated with rabbit antibodies to HA, NA (anti-HA MAb (11055-RM05), anti-NA MAb (11058-R001) and anti-NP PAb (11675-RP01) all from Sino Biological, China), 1 µg/mL in PBS (100 µl/well), and kept overnight at +4°C 0.5% BSA-PBS was used for post-coating. After washing with PBS, Pandemrix antigen suspension (100 µl/well) was added and incubated in the antibody coated wells. After washing, mouse monoclonal antibodies to HA, NA or NP (anti-HA MAb (11055-MM08), anti-NA Mab (11058-MM07) or anti- NP MAb (11675-MM03) (1 µg/ml in 0.5% BSA-PBS) were added. In the EIA for IgG-antibodies to Pandemrix derived NP, plasma samples were diluted 1∶300 in 0.5% bovine serum albumin (BSA)-PBS. The binding to antibodies was detected with alkaline phosphatase conjugated anti-human IgG-antibody or anti-mouse IgG-antibody (both from Jackson ImmunoResearch, USA). After adding the substrate the color reaction was read.

Polystyrene plates were coated with recombinant HA from H1N1 influenza A/California/07/2009 vaccine strain or NP from A/Puerto Rico/8/1934 (H1N1) in PBS (both from Sino Biological, China), 0.1 µg/well. 0.5% BSA-PBS was used for post-coating, washing buffer was either 0.5% BSA-PBS or 0.5% BSA-0.05% PBS-polysorbate 80, and plasma samples were diluted 1∶100 in 0.5% BSA-PBS or 0.5% BSA-0.05%PBS-polysorbate 80. Alkaline phosphatase conjugated anti-human IgG-antibody (Jackson ImmunoResearch, USA) or anti-mouse IgG antibody (Jackson ImmunoResearch, USA) was used as a conjugate.