Plant-derived extracellular vesicles (PDEVs) have attracted interest for potential use in cosmetics, aesthetic medicine ingredients, functional foods and drug delivery. Unlike animal cell-culture supernatants, plant tissues contain cell walls, polysaccharides, polyphenols, pigments and fibers. Inadequate pretreatment can cause membrane particles from broken cells and other impurities to co-enrich with vesicles. Isolation should therefore be designed around sample properties, target purity and downstream use rather than a fixed kit or single workflow.
A complete plant EV isolation and characterization workflow generally includes sample pretreatment, supernatant collection, isolation and purification, characterization, and quality control and delivery. Drawing on Lifeint’s reported experience of more than 20,000 EV-related projects and coverage of over 300 plant species, including more than 160 medicinal plants, this guide presents technical points, reference parameters and common pitfalls.
Three common plant-vesicle sample sources are:
For dense tissues such as roots and stems, low-temperature grinding may increase extractable particles, but the resulting sample can also contain membrane particles from broken cells. The source recommends stating this limitation in study design and reporting, rather than equating nanovesicles obtained by different preparation routes.
The source describes enzymatic digestion or culture as ways to obtain an EV-containing supernatant for subsequent isolation.
The source notes that plant EV content is often lower than that of animal-derived samples and recommends sufficient supernatant volume (≥50 mL) to obtain enough particles for downstream characterization and omics after purification.
This is a key step affecting EV purity, integrity and yield. Common methods are compared below for different sample volumes and purity requirements:
| Method | Principle | Reference price | Turnaround | Applications |
|---|---|---|---|---|
| Ultracentrifugation (standard) | Differential ultracentrifugation (UC) | 200 CNY/50 mL | 3 business days | Conventional samples; classic workflow |
| Ultracentrifugation (sucrose cushion) | Sucrose cushion to reduce protein impurities | 300 CNY/50 mL | 3 business days | Samples requiring improved purity |
| Ultracentrifugation (iodixanol gradient) | OptiPrep density-gradient centrifugation | 500 CNY/50 mL | 3 business days | High-purity applications; ≥6 samples per batch |
| Polymer precipitation (PEG) | PEG precipitation | 200 CNY/50 mL | 3 business days | Fast and low cost |
| Ultrasound nanofiltration (EXODUS) | Ultrasound nanofiltration chip | CNY 600/500 μL | 5 business days | Small, precious samples (≤1 mL) |
| Magnetic-bead capture (MSP membrane-sensing peptide) | Membrane-sensing peptide-conjugated beads | 200 CNY/sample | 3 business days | Rapid total EV enrichment (≤1.5 mL) |
| Large-volume isolation (EXO-DM) | Multistage membrane filtration + tangential-flow concentration + size-exclusion purification | CNY 1,500/L | 10 business days | Samples ≥3 L; industrial-scale applications |
For industrial demand in areas such as cosmetic, aesthetic medicine and food ingredients, the source describes Lifeint’s EXO-DM large-volume isolation equipment using tangential-flow filtration (TFF) and size-exclusion chromatography (SEC), with the following process:
Supernatant pretreatment → porous-membrane coarse filtration → ceramic-membrane fine filtration → disc dual-phase sterile filtration → ultra-high-pressure tangential-flow concentration → low-pressure high-throughput size-exclusion chromatography → iodixanol-cushion ultracentrifugation and concentration → purity and contaminant testing
The source reports that this process has been used for isolation and characterization across over 300 plant species and can be adjusted for downstream requirements (reference price: 1,500 CNY/L; sample volume ≥3 L).
After isolation, multiple measures can be combined to assess EV morphology, particle size, concentration and marker proteins:
| Assay | Method / platform | Reference price | Turnaround | Details |
|---|---|---|---|---|
| Electron-microscopy morphology | TEM negative staining | 200 CNY/sample | 3 business days | Six images by default: four fields at 1,000, 500, 200 and 100 nm |
| Particle size and concentration | Nano-flow cytometry (NanoFCM) | 200 CNY/sample | 3 business days | Recommended by default; single-particle analysis |
| Particle size and concentration | NTA(Zetaview) | 200 CNY/sample | 3 business days | Nanoparticle tracking analysis |
| Particle size and concentration | NTA(NS300) | 400 CNY/sample | 5 business days | High-resolution option |
| Protein quantification | BCA assay | 50 CNY/sample | 1 business day | Total protein concentration after lysis |
| Protein markers | Western Blot | 100 CNY/sample/marker | 3 business days | Six antibodies provided at no charge: CD9, CD63, CD81, TSG101, Syntenin1 and Calnexin |
| Protein markers | Nano-flow cytometry (NanoFCM) | 200 CNY/sample/marker | 3 business days | CD9, CD63 and CD81 flow antibodies provided at no charge |
Plant EV marker note: the source notes that classical tetraspanins such as CD9, CD63 and CD81 may be less abundant in plant-derived than mammalian EVs. Confirm antibody suitability for the species, particularly outside human, rat and mouse, and consider plant-specific markers such as PEN1 or TET8 where appropriate.
For material and industrial applications, the source suggests considering these additional quality measures:
| Quality measure | Method | Reference price | Turnaround |
|---|---|---|---|
| Purity assessment | Lysis method (comparison before and after treatment with triton-100) | 500 CNY/sample | 3 business days |
| Purity assessment | Membrane dye method (Evmembrane Green Stains) | 500 CNY/sample | Three business days (animal-derived samples only) |
| Zeta potential | NTA(Zetaview) | 300 CNY/sample | 3 business days |
| Osmolality | SMC 30D osmometer | 200 CNY/sample | 3 business days |
| Microbial testing | 24-hour culture in medium with shaking | 200 CNY/sample | 3 business days |
| Mycoplasma testing | qPCR | 200 CNY/sample | 3 business days |
| Endotoxin testing | Limulus amebocyte lysate assay | 200 CNY/sample | 3 business days |
| Heavy-metal testing | Atomic absorption spectroscopy (AAS) | 600 CNY/sample | Five business days (lead, cadmium, chromium, mercury and arsenic) |
Q1: How do isolation methods differ for plant and animal EVs?
A: The source identifies sample pretreatment as the main difference. It describes cellulase and pectinase digestion for plant cell walls (≤3 g tissue, 200 CNY/sample) and trypsin or collagenase for many animal samples, followed by shared isolation and purification steps.
Q2: What is the minimum sample amount?
A: The source estimates tens to hundreds of micrograms from 3 g of raw material in a small test and a few milligrams from 1 kg in batch extraction. Estimate input based on the EV amount needed downstream.
Q3: How long does isolation and characterization take?
A: The source lists approximately three business days for ultracentrifugation, particle sizing and TEM; about 20 business days with omics; and about ten business days for large-volume EXO-DM isolation (≥3 L).
Q4: Which plant species are supported?
A: Lifeint reports coverage of over 300 plant species, including more than 160 medicinal plants such as ginseng, astragalus, Panax notoginseng, aloe, Centella asiatica, tea, grape and loquat. Consult the species catalogue or contact the company.
Q5: Can the isolation process be customized?
A: The source says the EXO-DM large-volume workflow and quality controls can be adjusted for downstream needs, with customized plans quoted separately.
For full service pricing, see Service pricing; for transcriptomics, proteomics or metabolomics, see Omics analysis guide; to request services, visit Isolation and characterization services.
Last updated: 2026-08-12
*Prepared by Xiamen Lifeint Technology. Reference prices and timelines follow Pricing ; specific terms are subject to the agreement between the parties.*
Lifeint provides EV research and application services from sample preparation, isolation and characterization to omics, functional studies, modification and scale-up. It jointly established a biomedical technology centre with Xiamen University School of Pharmaceutical Sciences, built process-development laboratories to GMP standards, and contributed to an EV expert consensus. Its service records cover more than 300 plant species.
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