Key assay validation parameters tested included specificity, accuracy, precision, limit of detection and quantitation, specificity, linearity and range, and robustness.
By combining the approach of design of experiments (DOE) with subject matter expertise and experience, we were able to understand thoroughly how assay parameters affect the performance of the assay individually and interactively, identify the key assay parameters, define assay operating ranges and finally achieve a robust and sensitive cell-based assay for the detection of NAbs to Conatumumab.
Indirect immunofluorescence microscopy, scoring the formation of punctate intracellular vacuoles stained for LC3B, is a key assay to monitor autophagy and to understand its dynamics.
Third, a key assay for specificity is the ability of RNA derived from the corresponding human or zebrafish cDNA to prevent the LOF phenotype, when co-injected with the appropriate MO.
Overall, the system (plus the additional simple extensions to key functional assays) represents an important advancement towards the routine use of three-dimensional cultures for preclinical oncology drug development.
We strongly believe that T-µTP is a suitable model for the study of breast cancer and for drug screening assays following key parameters of clinical interest.
For this purpose, we focused on developing and validating nine assays for key drug resistance mutations in subtype B HIV-1 clinical specimens as a basis for later expansion to other virus mutations and subtypes.
Most importantly, the in vivo animal assays provided key evidences for the effect of miR-296-3p-ICAM-1 axis on M12/CTC escaping from NK cell lysis in blood vessels.
A key limitation of scratch assays is the question of whether they are reproducible since the scratch can be made with various types of instruments and varying degrees of pressure, and the assay can be performed on several different types of substrates.
As kinetics is the key indicator for assaying immunogenicity, this strategy would lack general applicability.
