Immunoassays include detection of binding of antibody-conjugated drugs to target proteins; quantitative ADC analysis; evaluation of stability of linkers and drugs; detection of binding to FcγR and evaluation of immunogenicity. These assays are important for understanding the potency, pharmacokinetics, and safety of antibody-conjugated drugs. Among them, enzyme-linked immunosorbent assays are often used to evaluate the concentration of antibody-drug conjugates and their binding to antigens.
ELISA types for antibody-drug conjugates analysis include total antibody ELISA, semi-homogeneous ELISA, competitive ELISA, and coupled antibody ELISA. The specific total antibody ELISA refers to coating a target antigen or an anti-idiotypic antibody on a well plate, and using an enzyme-specific anti-immunoglobulin secondary antibody to detect the binding of the ADC to the coated antigen. In the case of a semi-homogeneous ELISA, the capture antibody and the detection antibody form a complex with the ADC prior to immobilization to the well plate, and exhibit significant advantages in analyzing antibody-conjugated drugs of different drug loadings.
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