Are specifications or documents available for ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit?

Yes. The page provides ExoCoupl EC-01-02 EVs表面修饰生物正交偶联试剂盒说明书.pdf. Detailed specifications should be confirmed with the page or official documents.

What should be confirmed before using ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit?

Users should confirm sample type, experimental goal, starting volume or particle concentration, downstream assays, and whether supporting purification, characterization, or tracing workflows are required.

ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit

Overview ExoCoupl ® exosome surface modification bioorthogonal coupling kit is a kit developed by Echo biotech to achieve exosome surface modification based on bioorthogonal reactions.

Quick Facts

Exosome Engineering Modification
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Overview ExoCoupl ® exosome surface modification bioorthogonal coupling kit is a kit developed by Echo biotech to achieve exosome surface modification based on bioorthogonal reactions.

Overview

ExoCoupl ® exosome surface modification bioorthogonal coupling kit is a kit developed by Echo biotech to achieve exosome surface modification based on bioorthogonal reactions. First, active groups are added to the surface of exosomes through room temperature bioorthogonal reaction, and then through the reaction between the active group and the azide bond (N3), active ingredients such as small molecules/polypeptides/nucleic acids coupled with azide are covalently bound to the outer surface of exosomes to achieve surface modification of exosomes. The active ingredients of azide coupling are relatively convenient to synthesize and have no restrictions on electrical properties, molecular types, etc. After synthesis, they can be connected to the surface of exosomes using this kit. This coupling kit is easy to operate, requiring only 2 steps of co-incubation and 2 steps of purification. The exosome surface loading efficiency is as high as that of lipid anchor co-incubation, but the non-specific adsorption is lower.

Specifications

ProductName | CatalogNumber | Specification | Application: ExoCoupl® Exosome Surface Modification Bioorthogonal Conjugation Kit | EC-01-02 | 6T | Supports 6 bioorthogonal conjugation reactions and can process 500 μL × 6 = 3 mL of exosomes (recommended exosome concentration: >1E+11 particles/mL or 0.5–1 μg/μL)

Overview

Specifications

Applications

1. Exosome loading of active ingredients such as therapeutic small molecules, small nucleic acids, and peptides; 2. Surface loading of targeting peptides to improve the targeting of exosomes, such as cranial nerve targeting peptide RVG, ischemic myocardial targeting peptide IMTP, pan-cancer or inflammatory cell targeting peptide iRGD, etc.

Advantages

1. Easy to operate, only 2 steps of co-incubation and 2 steps of purification are required to complete exosome surface loading; 2. The exosome surface loading efficiency is high. For example, for targeting peptide exosome surface loading, the loading positive rate of this kit is comparable to that of lipid anchor co-incubation. 3. This kit uses a 2-step labeling method to load target molecules with lower non-specific adsorption and lower background noise. 4. This kit does not limit the type of target molecules that can be loaded. User-prepared N3-labeled small molecules, peptides, and nucleic acids can be used, and the synthesis is convenient. Of course, target molecules less than 4KD have higher loading efficiency. In order to facilitate the measurement of loading, it is recommended to bring N3 and FITC/FAM and other fluorescent groups at the same time.

Performance

First, synthesize N3 and FITC-labeled cranial nerve targeting peptide RVG (FITC-RVG-K-N3), and use this orthogonal coupling kit (Cat#EC-01-01) to process Echo biotech shelf product 293F exosomes (Cat#CTE-06) according to the instructions to modify the RVG targeting peptide on the exosome surface. At the same time, another portion of 293F exosomes (Cat#CTE-06) was used, and the RVG lipid-anchored targeting peptide (Cat#EA-06-1) was used to display the RVG targeting peptide carrying FAM fluorescence on the surface of the exosomes. 1. Loading rate comparison: Use NanoFCM to detect the positive rate of RVG peptide loading (the number of exosome particles loaded with RVG/the number of all exosome particles). As shown in the figure below, the results show: 1) Both methods can modify RVG peptides onto the exosome surface, and the positive loading rates are similar; 2) The background signal is lower when loaded with the orthogonal coupling kit. 2. The RVG-modified 293F exosomes obtained by the above two methods, the exosomes obtained by the lipid anchor method were named "lipid-anchored RVG-exosomes", and the exosomes obtained by the orthogonal coupling method were named "conjugated RVG-exosomes". RVG modification was not performed. The decorated exosomes were named "control 293F exosomes". After the above three groups of exosomes were labeled with red fluorescence, mouse brain neuroma cells Neuro2a cells and prostate cancer cells were treated respectively, and the cell flow cytometry was used to detect the uptake rate of exosomes by the cells. The results showed that the exosomes surface-modified with brain neuron-targeting peptide RVG obtained by the two methods could improve the uptake efficiency of exosomes by Neuro2a cells; while another group of prostate cancer cell controls that did not express RVG receptor protein showed no difference in uptake rate. Note: 1. Avoid repeated freezing and thawing. 2. Returns and exchanges are not accepted for non-product quality issues. 3. The verification data is for reference only and does not serve as a delivery standard. 4. Users need to synthesize drug molecules with fluorescent groups and azide bond coupling by themselves. IF there are difficulties in synthesis, please contact technical support to recommend a synthesis supplier; 5. It is recommended that the molecular weight MW of the target molecule should not be greater than 4 kD to obtain the best coupling effect. IF the molecular weight is large, the dosage can be appropriately increased in experimental step 5. The initial recommended dosage calculation method is MW*10/4000 μl (the concentration of N3-target molecule-FITC is 1 mg/ml). 6. The maximum loading volume of the mini column does not exceed 600 μl. 7. Please shake and mix the liquid solution before use, and avoid light when operating. 8. Shelf life: 1 year.

FAQ

1. How to calculate the loading capacity after using a multifunctional microplate reader to detect fluorescence? Use the synthesized N3-target molecule-FITC solution of known concentration (1 000 μg/ml) as a standard, add 20 μl to 180 μl of universal buffer and dilute 10 times to 100 μg/ml, then add 100 μl to 100 μl of universal buffer, dilute 2 times to 7 concentration gradient standards, and mix each one; the exosome final product sample is diluted about 2-10 times with universal buffer to Mix 100 μl; add 80 μl of each concentration standard and sample to a 96-well plate, and use a multifunctional microplate reader to detect the fluorescence intensity (excitation light 488nm, emission light 515nm-535nm). U According to the fluorescence value and corresponding concentration (μg/ml) of the standard substance, the microplate reader is used to obtain the standard curve: y=Ax+D U Fluorescence value of diluted exosome final product detected by microplate reader: B (EU) U Dilution factor of final exosome product: n U N3 - Molecular weight of target molecule-FITC: M (Da) U Total amount of exosome final product: BCA detection concentration is C (μg/ml) Then, the total load of lipid anchor targeting peptide per unit of exosomes (nmol/mg) = [(B-D)/A] *n/M/C*1000000 2. How to store loaded exosomes? For best results it is recommended to use fresh samples whenever possible, but due to limitations of experimental conditions it may not be possible to use fresh samples every time. In order to ensure the proportion of positive particles, it is recommended to store the sample at 4°C for 3 days and no more than 7 days at most, and to store it at -20°C for 2 weeks. Samples should be kept away from repeated freezing and thawing as much as possible (appropriate amounts can be aliquoted before freezing).

Manual Downloads

References

ExoCoupl ® kit cited literature: Lin H, Yin L, Liu W, et al. Muscle-Derived Small Extracellular Vesicles Mediate Exercise-Induced Cognitive Protection in Chronic Cerebral Hypoperfusion. Adv Sci (Weinh) . Published online April 24, 2025. doi:10.1002/advs.202410209 IF14.1 Zone 1 Literature download link: https://advanced.onlinelibrary.wiley.com/doi/10.1002/advs.202410209 Purpose: Use the ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit to modify the N3-FITC reference substance included in the kit onto the exosome surface according to the operating instructions to perform FITC fluorescent labeling modification on the exosomes. Then the tissue was obtained by intravenous injection, and the exosomes were traced with Ex Vivo after sectioning. Results: Fluorescein Isothiocyanate (FITC)+ sEVs can also be detected in the hippocampus (Figure S2C, Supporting Information).

Documents

ExoCoupl EC-01-02 EVs表面修饰生物正交偶联试剂盒说明书.pdf

FAQ

What is ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit?: ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit is an exosome research reagent under the Exosome Engineering Modification category. Overview ExoCoupl ® exosome surface modification bioorthogonal coupling kit is a kit developed by Echo biotech to achieve exosome surface modification based on bioorthogonal reactions.
What experiments is ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit used for?: ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit is suitable for exoCoupl ® exosome surface modification bioorthogonal coupling kit is a kit developed by Echo biotech to achieve exosome surface modification based on bioorthogonal reactions. First, active groups are added to the surface of exosomes through room temperature bioorthogonal reaction, and then through the reaction between the active group and the azide bond (N3), active ingredients such as small molecules/polypeptides/nucleic acids coupled with azide are covalently bound to the outer surface of exosomes to achieve surface modification of exosomes. The active ingredients of azide coupling are relatively convenient to synthesize and have no restrictions on electrical properties, molecular types, etc. After synthesis, they can be connected to the surface of exosomes using this kit.
Which product category does ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit belong to?: ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit belongs to the Exosome Engineering Modification category and can be evaluated together with related exosome reagents for culture, isolation, purification, engineering, characterization, or tracing workflows.
Are specifications or documents available for ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit?: Yes. The page provides ExoCoupl EC-01-02 EVs表面修饰生物正交偶联试剂盒说明书.pdf. Detailed specifications should be confirmed with the page or official documents.
What should be confirmed before using ExoCoupl ® Exosome Surface Modification Bioorthogonal Coupling Kit?: Users should confirm sample type, experimental goal, starting volume or particle concentration, downstream assays, and whether supporting purification, characterization, or tracing workflows are required.