Publisher Summary Studies of receptor binding generally employ highly radioactive ligand derivatives; however, knowledge of the biological activity of the radioactive derivative is essential for a complete interpretation of the binding data. This chapter describes the relation between biological activity and binding, physicochemical detection and assay of receptors, solubilization of membrane receptors, provides examples of membrane-localized receptors, affinity techniques, polyvalent hormone derivatives, and receptor studies, and theoretical mechanisms of hormone–receptor binding and action. The biological response toward many neurotransmitters and hormones, such as insulin, acetylcholine, and epinephrine is rapid both in onset and in the termination of action on removal of these agents; therefore, it may be anticipated that the initial hormone–receptor interaction will also be reversible. The chapter shows that progress has been made toward understanding the way by which hormones, via interactions with cell–surface receptors, modulate overall cellular function. This interaction is only the first in a presumably complicated series of protein–protein interactions within the plane of the cell membrane; the mechanism, whereby the ultimate signal is transmitted to the internal cellular milieu is undoubtedly an equally intricate process. The chapter is considered to be useful in delineating the challenge that the elucidation of these processes presents and in providing a basis and incentive for more discoveries in this field.

Membrane receptors and hormone action.

Key aspects of analytical method validation and linearity evaluation.

Studies comparing a new method with an established method, to assess whether the new measurements are comparable with existing ones, are frequently conducted in clinical pathology laboratories. Assessment usually involves statistical analysis of paired results from the 2 methods to objectively investigate sources of analytical error (total, random, and systematic). In this review article, the types of errors that can be assessed in performing this task are described, and a general protocol for comparison of quantitative methods is recommended. The typical protocol has 9 steps: 1) state the purpose of the experiment, 2) establish a theoretical basis for the method comparison experiment, 3) become familiar with the new method, 4) obtain estimates of random error for both methods, 5) estimate the number of samples to be included in the method comparison experiment, 6) define acceptable difference between the 2 methods, 7) measure the patient samples, 8) analyze the data, and 9) judge acceptability. The protocol includes the essential investigations and decisions needed to objectively assess the overall analytical performance of a new method compared to a reference or established method. The choice of statistical methods and recommendations of decision criteria within the stages are discussed. Use of the protocol for decision-making is exemplified by the comparison of 2 methods for measuring alanine aminotransferase activity in serum from dogs. Finally, a protocol for comparing simpler semiquantitative methods with established methods that measure on a continuous scale is suggested.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1939-165X.2006.tb00131.x

Method comparison in the clinical laboratory

We have studied the usefulness of common statistical tests as applied to method comparison studies. We simulated different types of errors in test sets of data to determine the sensitivity of different statistical parameters. Least-squares parameters (slope of least-squares line, its y intercept, and the standard error of estimate in the y direction) provide specific estimates of proportional, constant, and random errors, but comparison data must be presented graphically to detect limitations caused by nonlinearity and errant points. t -test parameters ( bias, standard deviation of difference) provide estimates of constant and random errors, but only when proportional error is absent. Least-squares analysis can estimate proportional error and should be considered a prerequisite to t -test analysis. The correlation coefficient ( r ) is sensitive only to random error, but is not easily interpreted. Values for r, t , and F are not useful in making decisions on the acceptability of performance. These decisions should be judgments on the errors that are tolerable. Statistical tests can be applied in a manner that provides specific estimates of these errors

Use and Interpretation of Common Statistical Tests in Method Comparison Studies

In the clinical laboratory setting, interferences can be a significant source of laboratory errors with potential to cause serious harm for the patient. After hemolysis, lipemia is the most frequent endogenous interference that can influence results of various laboratory methods by several mechanisms. The most common preanalytical cause of lipemic samples is inadequate time of blood sampling after the meal or parenteral administration of synthetic lipid emulsions. Although the best way of detecting the degree of lipemia is measuring lipemic index on analytical platforms, laboratory experts should be aware of its problems, like false positive results and lack of standardization between manufacturers. Unlike for other interferences, lipemia can be removed and measurement can be done in a clear sample. However, a protocol for removing lipids from the sample has to be chosen carefully, since it is dependent on the analytes that have to be determined. Investigation of lipemia interference is an obligation of manufacturers of laboratory reagents; however, several literature findings report lack of verification of the declared data. Moreover, the acceptance criteria currently used by the most manufacturers are not based on biological variation and need to be revised. Written procedures for detection of lipemia, removing lipemia interference and reporting results from lipemic samples should be available to laboratory staff in order to standardize the procedure, reduce errors and increase patient safety.

/pdf/lipemia-causes-interference-mechanisms-detection-and-4exi17yri4.pdf

Lipemia: causes, interference mechanisms, detection and management

We describe an approach for formulating criteria that can be used to judge whether an analytical method has acceptable precision and accuracy. We derive criteria for several experiments that are commonly used in method-evaluation studies: precision or replicates, recovery, interference, and comparison of patient values between the new method and a proven method. These criteria are based on the medical usefulness of the test results, thus the acceptability of the method is judged with respect to the clinical requirements.

Criteria for Judging Precision and Accuracy in Method Development and Evaluation

An unbiased method for estimating binding parameters in a non-cooperative binding process.

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Criteria for Judging Precision and Accuracy in Method Development and Evaluation

An unbiased method for estimating binding parameters in a non-cooperative binding process.