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Determination of antigen-antibody binding affinity and apparent association constant by Langmuir plot. Antigen used in this assay were Gly-HSA. Immune complexes were prepared by incubating 100 µg of IgG from IV-S (-●-) and IV (-▲-) subjects (A) and IgG from III-S (-●-) and III (-▲-) subjects (B) with varying amount of antigen (0–100 mg) in an assay volume of 100 µL for 2 h at room temperature and overnight at 4 °C.
