Exosome Biotinylation Service

Exosome biotinylation is a technique that covalently labels the surface proteins or lipid structures of exosomes with biotin molecules through chemical or enzymatic methods. Biotin, as a small molecular "tag," exhibits an extremely high affinity (Kd ≈ 1e-15 M) for streptavidin, enabling highly specific binding. This feature allows for efficient capture, purification, or visualization of biotin-labeled exosomes. The advantages of exosome biotinylation:

1. Simplified Isolation and Purification: Biotinylated exosomes can be rapidly enriched using streptavidin-coated magnetic beads, microfluidic chips, or solid-phase carriers, minimizing sample loss and nonspecific adsorption.

2. Enhanced Detection Sensitivity: The biotin-streptavidin system can be coupled with enzyme-linked colorimetric assays, fluorescent probes, or quantum dots, significantly improving the detection limits of low-abundance exosomal surface proteins.

3. Multifunctional Research Applications: Labeled exosomes can be directly used for interaction protein identification, cellular uptake tracking, and multi-marker analysis, facilitating both fundamental research and clinical translation.

 

MtoZ Biolabs provides end-to-end Exosome Biotinylation Service covering exosome isolation, site-directed biotinylation, and functional validation, dedicated to solving key challenges in exosome research such as low labeling efficiency, high sample loss, and insufficient data reliability. Through optimized biotinylation systems and stringent quality control, we ensure authentic expression and biological activity of exosomal surface markers. Our exosome biotinylation service features:

1. Flexible Compatibility: Adaptable to various sample types including serum, plasma, and cell culture supernatants.

2. High Labeling Efficiency: Targeting exosomal lipid membranes or specific surface proteins (e.g., CD63, CD9), with labeling efficiency >90%.

3. Non-destructive Processing: Utilizing non-disruptive biotinylation reagents to maintain exosome membrane integrity and preserve RNA/protein content stability.

 

Analysis Workflow

1. Exosome Isolation and Purification

• Ultracentrifugation (UC) combined with Size Exclusion Chromatography (SEC) to remove lipoproteins and free proteins.
• Dual validation by Transmission Electron Microscopy (TEM) and Nanoparticle Tracking Analysis (NTA), ensuring exosome purity (≥1e8 particles/mL) and size range (50-150 nm).

 

2. Optimized Biotinylation Labeling (Strategy Tailored to Application)

• Membrane Lipid Labeling: Using water-soluble biotinylation reagents (e.g., Sulfo-NHS-LC-Biotin) for broad-spectrum binding to amino groups on exosomal membranes.
• Targeted Protein Labeling: Employing biotinylated antibody conjugates for specific labeling of surface markers like CD63, EGFR.
• Optimized reaction conditions (pH, temperature, incubation time) to minimize aggregation and denaturation risks.

 

3. Post-labeling Validation and Quality Control

• Labeling Efficiency Assessment: Quantification of streptavidin binding capacity via flow cytometry or ELISA.
• Functional Verification: Co-culture assays to validate the cellular uptake activity of biotinylated exosomes (e.g., fluorescence tracking).

 



Lee, BR. et al. J Extracell Vesicles. 2022.

Figure 1. An in situ Strategy for Labelling Exosome Proteins by Using the Engineered Ascorbate Peroxidase APEX

 

Why Choose MtoZ Biolabs?

 

Table 1.  The Advantages of Exosome Biotinylation Solutions at MtoZ Biolabs

Aspect	Traditional Methods	MtoZ Biolabs Solutions
Labeling Efficiency	40–60% via chemical conjugation	>90% (optimized reaction system)
Sample Compatibility	Requires highly concentrated exosomes	Compatible with low-abundance samples (≥1e7 particles/mL)
Marker Preservation	Risk of membrane protein denaturation	Gentle reagents preserve protein conformation and function
Detection Sensitivity	Limited to WB/ELISA	Supports single exosome flow cytometry and proximity barcoding
Turnaround Time	3–5 days (outsourced in steps)	Full-process one-stop service, ≤72 hours

 

Applications

1. Cancer Liquid Biopsy

• Biomarker Discovery: Capturing biotinylated exosomes for mass spectrometry-based identification of tumor-specific surface proteins.
• Therapeutic Monitoring: Dynamic tracking of PD-L1 expression on exosomes from chemotherapy patients to assess immunotherapy response.

 

2. Drug Delivery and Vaccine Development

• Targeted Modification: Loading targeting molecules or drugs via the biotin-streptavidin system to construct intelligent delivery vectors.
• In Vivo Distribution Tracking: Fluorescent imaging of biotinylated exosomes to monitor their organ-specific accumulation in animal models.

 

3. Infection and Immunity Studies

Pathogen Interaction Analysis: Labeling host exosomal receptors to study their binding mechanisms with viral or bacterial proteins.

 

Deliverables

1. Exosome Isolation and Standardized Pre-processing (≥1 mL sample, NTA particle size distribution report).

2. Biotinylation Strategy Development: Including reagent screening, concentration gradient testing, and stability validation.

3. Quality Control Report: Labeling efficiency (flow cytometry), exosome integrity (TEM), and functional data (e.g., cellular uptake assay images).

4. Technical Consultation: Support for downstream applications such as single exosome multiplex detection.

 

The biological functions and clinical values of exosomes are increasingly recognized, and precise labeling technologies are critical to realizing their translational potential. MtoZ Biolabs, with its robust experimental platform, rigorous quality control, and personalized support, is committed to providing high-reliability exosome biotinylation solutions for researchers worldwide. Whether for fundamental studies of exosome biology or the development of novel diagnostic tools, we are your trusted partner. Contact us today to empower your exosome research with precise biotinylation!