EV / Exosome Analytical Services

Exosomes / Extracellular Vesicles (EVs) are an emerging class of biologics that play a crucial role in intercellular communication and hold tremendous promise for therapeutic and diagnostic use. A key challenge in the clinical translation of exosome-based therapies is the implementation of robust analytical methods for characterization. The size, heterogeneity, and complex composition of exosomes creates challenges for the use of conventional analytical tools and strategies, especially for critical quality attributes such as purity and potency.

RoosterBio has developed an extensive panel of analytical methods to support process development and manufacturing of exosomes, which we offer to our customers as either a stand-alone service or as an integrated component of a broader process development program. Our standard exosome analytics are bundled into a Base Package and Plus Package, along with additional supplemental assays to meet a broad range of customer needs, regardless of their cell type or production process.

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We offer two EV / Exosome Analytics Packages and Supplemental Assays specially designed for starting and advanced programs alike:

Assay Assay Result EV / Exosome Base Package EV / Exosome Plus Package Supplemental Assay
Nanoparticle Tracking Analysis (NTA) Particle Number, Particle Size
EV / Exosome Purity % EV by Particle Concentration
BCA Quantitation Total Protein
Surface Marker Expression Marker Expression
Cytosolic Marker Expression Alix Expression
miRNA Quanitation miRNA Content
miRNA Identity Analysis miRNA Identity
Lipid Content Total Lipid Content
Albumin Contamination Analysis Process Related Contaminant
CD63 Quantitation Quantified Marker Expression
Scratch Assay Wound Healing

Nanoparticle Tracking Analysis (NTA)

Nanoparticle Tracking Analysis is one of the most widely used methods for characterizing EV identity. This assay utilizes light scattering to quantify the size distribution and concentration of particles in a sample which are two key identity components of EV characterization. Results are provided as a concentration in particles per mL and a size distribution graph of EV diameters.

EV / Exosome Purity

This nanoflow cytometry assay for EVs measures the purity of your samples as a percent of particle population with membrane-bound lipids. As a result, these results will inform you what percentage of your particle concentration is made up of EVs. Additional assays can assess tetraspanin make-up of the particle population and custom target readouts for your target of interest upon request.

BCA Quantitation

This BCA ELISA is a widely used identity and content tool to measure total protein content of an EV sample. Quantitation of protein content is a useful metric for process optimization as well as characterization of a purified sample. Results are provided as total protein per mL and microBCA assays are available for samples beneath the range of detection.

Surface Marker Expression

CD9, CD63, and CD81 are tetraspanin proteins widely expressed in different cell types and considered identity markers of EVs. This western blot assay provides a sensitive, semi-quantitative readout via image and densiometric conversion for all three tetraspanins in sample preparations.

Cytosolic Marker Expression

This identity assay is a quantitative Alix ELISA which measures a MISEV-recognized cytosolic marker in EV preparations. Results are provided as Alix protein per mL.

miRNA Quantitation

Bioanalyzer quantitation of miRNA content, as well as miRNA size distribution, provides a critical content readout for understanding your EVs. Results are provided as miRNA concentration (miRNA per µL), gel view of miRNA migration, and electropherogram of size and signal intensity. This assay can also quantify total RNA should it be preferred.

miRNA Identity Analysis

Quantitatively identify specific miRNAs present in your EV samples. This real-time qPCR analysis provides 8 readouts across several functional categories as well as a minimum of two reference readouts. Additional readouts can be customized to your target of interest upon request.

Lipid Content

This plate reader-based fluorescence identity and content assay provides a quantitative measurement of total membrane-bound lipid content per mL.

Albumin Contamination Analysis

This quantitative ELISA measures human albumin which is a common media component and contaminant of EV preparations. Step-wise removal of albumin can be an informative analytical readout as well as a purity indicator for “final product” EV samples. Results of this purity assay are reported as albumin protein per mL.

CD63 Quantitation

Used for process development, this identity assay is a quantitative ELISA which measures a MISEV-recognized surface marker in EV preparations. Results are provided as CD63 protein per mL.