Today with the unprecedented Covid-19 pandemic, several researchers are looking at multiplexing options to study the cytokine storm. Cytokine storms are pro-inflammatory cytokines observed at elevated levels in severe Covid-19 patients. Hence, understanding the cytokine storm is necessary to predict the severity of Covid-19 cases. Several Luminex partners have already established multiplex assays for research use.
Luminex multiplex assays are solid-phase bead-based sandwich immunoassays designed to work with the Luminex 100/200 instrument. The spectral properties of these distinct bead regions can be used to detect up to 100 different target analytes in a single study sample. Such capabilities have made Luminex immunoassay the most widely employed multiplexing technique in drug development and discovery. However, what is the drug development process? The drug development process aims at identifying and assessing drug compounds that can be formulated into drug products against specific diseases and medical conditions.
Several Luminex ELISA assay kits are available for research, so selecting the most suitable Luminex multiplex assay can become difficult.. Understanding the strengths and limitations of each assay kit can help researchers make informed decisions. Hence here, we present seven crucial considerations for determining an ideal assay for research needs.
Dynamic Assay Range
The dynamic range must include the expected values of the study samples. In the case of endogenous samples, the dynamic range must cover lower concentrations. Whereas for disease state models, a dynamic range covering higher concentrations is suitable.. A large dynamic range works best when evaluating both types of samples
As endogenous analytes can be lower in quantity, the assay must be sensitive enough to detect low levels of endogenous analytes in study samples.
When choosing a Luminex multiplex assay kit, study the accuracy data from the vendor-provided sheet. This sheet is present in all assay kits. To evaluate the accuracy, the 80-120% range is the industry standard for spike-and-recovery experiments.
Inter and Intra-Assay Precision
Whether samples are analyzed on a single plate or multiple plates, the generated data must be precise and accurate. This necessity is particularly crucial when evaluating different treatment groups. Any observed variation should only be because of treatment effects and not due to variability in the assay.
Understanding and Evaluating lot-to-lot Variability
While performing long-term studies with many study samples, lot-to-lot variability is of utmost importance to ensure accurate, reliable, and consistent results. Particularly in preclinical research, where several compounds are tested over months or years, assessing a large number of study samples will exceed a single assay lot.
Determining Percent Cross-Reactivity Among Analytes
In the presence of multiple analytes in a single assay, cross-reactivity between the detection or capture reagents can generate inaccurate results. The vendor generally provides information about cross-reactivity. Researchers should not use kits with high cross-reactivity for analysis.
Accounting Matrix Effects
Bioanalytical studies involve assessing samples in several matrices, including serum, plasma, tissue homogenates, and cell culture media. All of these matrices can impact the performance of Luminex multiplex assays. Researchers must ensure that the assay kit provides buffers to prevent matrix effects or provide solutions for mitigating them.