Intra-assay precision was determined by analyzing 16 replicates of two serum controls in a single assay.

*In 5% cynomolgus monkey serum
**In 10% cynomolgus monkey serum
Inter-assay precision was determined by analyzing replicates of two serum controls in three separate assays on different days.

*In 5% cynomolgus monkey serum
**In 10% cynomolgus monkey serum

*O.D. + (1.64 x S.D.)
The lower limit of quantitation (LLOQ) is defined as the lowest standard concentration in which O.D. - (1.64 x S.D.) is higher than the blank value of O.D. + (1.64 x S.D.). The LLOQ is 7.8 ng/ml.
Matrix Validation:The Therapeutic Human IgG ELISA Kit, including the standard, does not contain serum of any species. The IgG4 standards have been analyzed in this assay in the presence of 1%, 5%, and 10% cynomolgus monkey and rat serum with no significant change in the characteristics of the assay (see Figure 5). When analyzing samples containing greater than 5% serum, we recommend diluting the standards in an equivalent serum concentration.

Spike Recovery:Cynomolgus monkey serum was spiked with different amounts of human IgG4, serially diluted with Assay Buffer (1X) and evaluated using the Human Therapeutic IgG4 ELISA kit. The error bars represent standard deviations obtained from multiple dilutions of the same sample.
