In Vitro ELISA and Cell-Based Assays Confirm the Low Immunogenicity of VNAR Therapeutic Constructs in a Mouse Model of Human RA: An Encouraging Milestone to Further Clinical Drug Development

Figure 2

Direct and indirect-binding ELISA for detecting functional serum anti-TNF-α drugs and nonneutralising mouse ADA. (a) Direct-binding ELISA of serum samples to 2 μg/mL hTNF-α coated wells, with functional drug binding determined using an anti-human IgG antibody- (Fc specific) peroxidase. (b) Indirect-binding ELISA capturing nonneutralising mouse ADA to anti-hTNF-α drugs. ELISA wells are coated with 2 μg/mL hTNF-α, and nonneutralising mouse ADA was detected using an anti-mouse IgG antibody- (whole molecule) peroxidase. All serum samples were randomly pooled and diluted at 1 : 20 in sterile PBS, pH 7.4. The results shown are the ( with two replicates per experiment).

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