Sample Replication Effects on Statistical Power in Multiplexed Immunoassay Analysis

It is known that by increasing replication via expanding the number of biological or technical replicates analysed, increases overall experimental precision and statistical power. The current and common immunoassay experimental practices, such as with Luminex xMap® technology, is that all samples are analysed in replicate. For most plate or array based immunoassay experiments this design of replication reduces by a half the actual numbers of biological replicates that can be analysed. Here it’s shown, in theory, that for common types of immunoassay experiments such as for screening biological samples for the concentration of analytes, that by leaving out the technical replicates doesn’t overly effect the expected value of the experiment’s assay variances, even though the number of assays have been halved. It is also shown that when the technical replicate assays are replaced by unique biological replicate assays, thereby keeping the number of assays the same, that the expected experimental variances coming from the biological replicates are decreased by 50%. Because the practice of technical replication in immunoassay analysis is current this report makes it explicit for plate and array based assay formats, that by using technical replicates over the option of biological replicates reduces overall experimental precision and efficiency of statistical analysis.