Statistical characterization of the random errors in the radioimmunoassay dose--response variable.

We have developed practical methods for evaluating the magnitude of the random errors in radioimmunoassay dose--response variables, and the relationship between this error and position on the dose--response curve. This is important: to obtain appropriate weights for each point on the dose--response curve when utilizing least-squares curve-fitting methods; to evaluate whether the standards and the unknowns are subject to error of the same magnitude; for quality-control purposes; and to study the sources of errors in radioimmunoassay. Both standards and unknowns in radioimmunoassays for cAMP and cGMP were analyzed in triplicate. The same mean (Y), sample standard deviation, sy, and variance (2-y) of the response variable were calculated for each dose level. The relationship between s 2-y and y was calculated utilizing several models. Results for standards and unknowns from several assays were pooled, and a curve smoothing procedure was used to minimize random sampling errors. This pooling increased the reliability of the analysis, and confirmed the presence of the theoretically predicted nonuniformity of variance. Thus, the calculation of results from these radioimmunoassays should utilize a weighted least-squares curve-fitting program. These analyses have been computerized, and can be used as a "pre-processor" for programs for routine analysis of results of radioimmunoassay.

[1]  A. Gilman A Protein BindingAssayforAdenosine 3':5'-Cyclic Monophosphate , 1970 .

[2]  C W Parker,et al.  Radioimmunoassay for cyclic nucleotides. I. Preparation of antibodies and iodinated cyclic nucleotides. , 1972, The Journal of biological chemistry.

[3]  D Rodbard,et al.  Computer analysis of radioligand assay and radioimmunoassay data. , 1970, Acta endocrinologica. Supplementum.

[4]  Curtis L. Meinert,et al.  The biometry of an isotope displacement immunologic microassay , 1968 .

[5]  M. Serio,et al.  Mathematical Analysis of the Results of Competitive Binding Methods , 1973 .

[6]  D Rodbard,et al.  Rapid calculation of radioimmunoassay results. , 1969, The Journal of laboratory and clinical medicine.

[7]  S. Wold,et al.  Evaluation of radio-isotope data in steroid assays based on competitive protein binding. , 1972, Clinica chimica acta; international journal of clinical chemistry.

[8]  S R Vivian,et al.  Classic bioassay statistical procedures applied to radioimmunoassay of bovine thyrotropin, growth hormone and prolactin. , 1971, The Journal of clinical endocrinology and metabolism.

[9]  A R Midgley,et al.  Principles for the assessment of the reliability of radioimmunoassay methods (precision, accuracy, sensitivity, specificity). , 1969, Acta endocrinologica. Supplementum.

[10]  D Rodbard,et al.  Statistical quality control and routine data processing for radioimmunoassays and immunoradiometric assays. , 1974, Clinical chemistry.

[11]  R. Rubin,et al.  Optimal statistical design of radioimmunoassays and competitive protein-binding assays. , 1975, Endocrinology.

[12]  J. Jacquez,et al.  Sampling Experiments on the Estimation of Parameters in Heteroscedastic Linear Regression , 1973 .

[13]  W G Duddleson,et al.  Computer program sequence for analysis and summary of radioimmunoassay data. , 1972, Computers and biomedical research, an international journal.

[14]  M. Healy,et al.  Statistical analysis of radioimmunoassay data. , 1972, The Biochemical journal.

[15]  D. Rodbard,et al.  A MODEL FOR PREDICTION OF CONFIDENCE LIMITS IN RADIOIMMUNOASSAYS AND COMPETITIVE PROTEIN BINDING ASSAYS. , 1970 .

[16]  G C Rennie,et al.  A generalized computer program for the treatment of data from competitive protein-binding assays including radioimmunoassays. , 1972, The Journal of laboratory and clinical medicine.

[17]  F. Greenwood,et al.  A radio-immunoelectrophoretic assay for human growth hormone. , 1964, The Biochemical journal.