Figure 6.

Characterization of rhesus immunoglobulin G (IgG) subclass composition and detection reagents. (A,B) Multiplex subclassing assays. (A) Recombinant monoclonal rhesus macaque IgG [Macaca mulatta (MM) IgG1–4] and pooled purified polyclonal rhesus macaque plasma IgG (MM pIgG) were titrated for binding to microspheres conjugated with anti-MM, subclass-specific antibodies (anti-MM IgG1, anti-MM IgG2, anti-MM IgG3), and bound test antibody detected by a pan anti-rhesus IgG detection antibody. (B) In a separate experiment, antibody purified from serum samples from 36 naïve MM (black) was likewise evaluated to detect the presence of IgG1, IgG2, and IgG3 subclasses. (C,D). ELISA subclassing assays. (C) Sensitivity and specificity of MM subclass detection reagents over a titration range when recombinant, CD8α-specific MM IgG subclass antibody test samples were bound to CD8α-conjugated ELISA plates. (D) The composition of MM serum IgG in a pooled sample was evaluated via an ELISA in which either non-specifically adhered serum IgG or recombinant MM IgG subclasses as controls were detected using a dilution series of the anti-subclass detection reagents.