METHODS Special Issue: Multiplexing Protocol for Evaluating Biological Interactions with Nanoparticles

A new multiplexing protocol offers guidelines for measuring immunomodulatory effects of nanoparticles used in clinical applications

METHODS Special Issue: Multiplexing Protocol for Evaluating Biological Interactions with Nanoparticles

Nanoparticles have the potential to deliver more targeted therapies to the exact location in the body where they’re most needed. To do this, it is essential to understand how these foreign particles interact with a patient’s body. A recent publication in the Methods journal is a big step forward, offering a protocol for evaluating the body’s immune response to nanotechnology.

Scientists from Texas A&M University and Bio-Rad Laboratories designed parameters for evaluating the immunomodulatory effects of nano- and microparticles using multiplex biomarker analysis. To detect biomarkers of interest, the researchers utilized xMAP® Technology from Luminex.

Considerations for monitoring the immunomodulatory effects of nanoparticles

According to the paper, the authors embarked on this project because “understanding the interactions of [particulate] materials with biological systems is crucial for the design of clinically-viable biomaterials,” they write. “The type and composition of nanomaterials plays a pivotal role in modulating the immune system, which may compromise therapeutic outcomes and can be life-threatening, [particularly] when the immunomodulation is unintentional.”

Multiplex techniques that allow for the analysis of as many as 500 biomarkers from a single sample would help address the need to characterize the effects of these particles, but certain challenges remain. For instance, the researchers note that “the adsorption of biomarkers on surfaces or within internal structures of nano- or microparticles has been explored to a lesser extent, although it can lead to biased conclusions and data misinterpretation.”

An empirical way to multiplex for large-scale biomarker analysis

With this protocol, the scientists aim to help other research teams looking to use multiplex technology to measure immunomodulatory effects of microparticles or nanoparticles by standardizing the techniques for both experiment preparation and analysis. The technique described is geared towards running 24 samples in triplicate on a 96-well plate, with appropriate standards and controls. The paper also includes precautions to help avoid artifacts, and to help with accurate data interpretation.

“The experimental details and precautions described [in the paper] allow for precise evaluation of biomarker expression in response to particulate materials, pharmaceuticals, and medical devices, and should be taken into careful consideration in pre-clinical and clinical studies, and other processes that involve the use of multiplexing techniques.”

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METHODS Special Issue: Multiplexing Protocol for Evaluating Biological Interactions with Nanoparticles