Calibration and LOD/LOQ estimation of a chemiluminescent hybridization assay for residual DNA in recombinant protein drugs expressed in E. coli using a four-parameter logistic model.
Calibration is the process of fitting a model based on reference data points (x, y), then using the model to estimate an unknown x based on a new measured response, y. In DNA assay, x is the concentration, and y is the measured signal volume. A four-parameter logistic model was used frequently for calibration of immunoassay when the response is optical density for enzyme-linked immunosorbent assay (ELISA) or adjusted radioactivity count for radioimmunoassay (RIA). Here, it is shown that the same model or a linearized version of the curve are equally useful for the calibration of a chemiluminescent hybridization assay for residual DNA in recombinant protein drugs and calculation of performance measures of the assay.
Lee
K R
KR
Statistical Sciences Department, SmithKline Beecham Pharmaceuticals R&D, King of Prussia, PA 19406, USA.
Dipaolo
B
B
Ji
X
X
D002138:Calibration; D003001:Cloning, Molecular / Q000379:methods*; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis* / Q000737:chemistry; D004274:DNA, Recombinant / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D008163:Luminescent Measurements; D008954:Models, Biological; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011863:Radioimmunoassay / Q000379:methods; D012015:Reference Standards; D015203:Reproducibility of Results
D015342:DNA Probes; D004269:DNA, Bacterial; D004274:DNA, Recombinant