Multiple analyte profiling (MAP) index as a powerful diagnostic and therapeutic monitoring tool.

A robust data mining algorithm is presented as a critical solution to the challenge of managing intensive data generated from the recently developed multiplexing techniques, which allow simultaneous detection of up to 500 biomarkers in a few microliters of a single sample. Furthermore, detailed methodology is provided for exploiting the new algorithm along with examples for description of the first application as a powerful diagnostic and therapeutic monitoring tool in the management of breast cancer, as a disease model.

Method to quantify cytokines and chemokines in mouse brain tissue using Bio-Plex multiplex immunoassays.

This protocol describes how to prepare mouse brain tissue for quantification of multiple inflammatory mediators using a multiplex bead-based immunoassay. It is important to have methods that allow quantification of multiple analytes from small amounts of tissue. Bio-Plex is a Luminex xMAP-based multiplex bead-based immunoassay technology that permits simultaneous analysis of up to 100 analytes from a single tissue sample. This assay has been used extensively to investigate analytes in plasma and serum samples as well as cultured and primary cells. Here, we describe a method for simultaneous analysis of 33 different inflammatory cytokines and chemokines from mouse brain tissue using the Bio-Plex Pro Mouse Chemokine Panel 33-Plex.

Multiplexing techniques for measurement of the immunomodulatory effects of particulate materials: Precautions when testing micro- and nano-particles.

Particulate materials at nano- and micro-scales have widespread pharmaceutical and medical applications. Understanding the interactions of these materials with biological systems is crucial for the design of clinically-viable biomaterials of high safety profiles. Immunomodulatory effects of particulate materials can be studied via multiplexing techniques that are capable of measuring up to 500 biomarkers in a few microliters of biological samples. However, there are several challenges towards the use of multiplexing techniques for testing the ability of nanomaterials to induce the release of various biomarkers. As one of the potential challenges, the adsorption of biomarkers on surfaces or within internal structures of nano- or micro-particles has been explored to a lesser extent, although it can lead to biased conclusions and data misinterpretation. Herein, we provide technical details on the use of multiplexing techniques for the evaluation of immunomodulatory effects of nanoparticulates. The same principles can also be applied for the assessment of microparticles. Importantly, precautions to avoid artifacts and data misinterpretation, due to interactions between particles and biomarkers, are provided.