Comprehensive Analysis of Pricella® Four Exosome Isolation Kits

Exosomes, as crucial mediators of intercellular comovery for downstream functional analysis, omics detection, and disease diagnosis. Traditional exosome extraction methods often face challenges such as high equipment requirements and complex workflows. To address these issues, Pricella® has developed a range of isolation solutions tailored to different sample types and research needs. In the previous Cell Culture Academy, we reviewed common exosome isolation methods. This time, we will focus on the four exosome isolation kits launched by Pricella®, providing a detailed analysis of their core principles, workflow, precautions, and advantages to help you choose the most suitable product.

I. Micro Exosome Isolation Kit

	Catalog No.: P-CA-501   Kit Components:   - Exosome Beads   - Solution A   - Elution Buffer

Principle Overview:

This kit utilizes proprietary homogenous liquid magnetic beads (Exosome Beads) that specifically bind to exosomes while excluding impurities. The magnetic separation technology enables high-purity and high-recovery isolation of exosomes.

Workflow:

1. Sample Pretreatment: Perform differential centrifugation (300×g, 5 min → 2000×g, 10 min → 14000×g, 30 min) to gradually remove cells, debris, and large particles.

2. Exosome Enrichment: Add the pretreated sample to the magnetic beads, incubate on a shaker at room temperature for 30 minutes.

3. Washing: Place on a magnetic rack, let it stand for 30 seconds, discard the supernatant, and wash twice with Solution A, retaining the magnetic beads.

4. Exosome Elution: Add Elution Buffer, incubate on a shaker at room temperature for 30 minutes. Place on the magnetic rack, let it stand for 30 seconds, and transfer the supernatant to a new centrifuge tube. The supernatant contains the isolated exosomes.

Precautions:

• In some samples, beads may aggregate when binding to exosomes; however, this does not affect the separation effect and does not require treatment.
• Isolated exosomes should not be stored in Elution Buffer for extended periods. For long-term storage, use ultrafiltration to replace the solution with PBS.

Advantages:

• High purity and integrity of isolated exosomes.
• Ideal for isolating exosomes from small or precious samples.
• Simple operation, requiring only a standard magnetic rack.
• High throughput, capable of processing multiple samples simultaneously, compatible with automated operation.

II. Universal Exosome Isolation Kit

	Catalog No.: P-CA-502/P-CA-503  Kit Components:  - Exosome Purification Column  - Column Adapter  Note: P-CA-502 and P-CA-503 differ only in product specifications and sample loading volume; the operation is identical.

Principle Overview:

Based on Size Exclusion Chromatography (SEC), the Exosome Purification Column is packed with porous gel beads that allow for the size-based separation of exosomes from other particles during PBS buffer flow.

Workflow:

1. Sample Pretreatment: Perform differential centrifugation (300×g, 5 min → 2000×g, 10 min → 14000×g, 30 min) to gradually remove cells, debris, and large particles.

2. Column Preparation: Equilibrate the purification column at room temperature for 30 minutes, check and remove any air, and balance the column with PBS.

3. Exosome Isolation: Load the pretreated sample into the column, add 0.5 mL PBS in batches, and collect the flow-through. Collect fractions 4-8 (P-CA-502) or 8-12 (P-CA-503) as isolated exosomes.

4. Column Maintenance: Wash with PBS and 20% ethanol, seal the column with 20% ethanol, and store at 4℃.

Precautions:

• PBS and 20% ethanol should be freshly prepared, filtered through a 0.2 μm membrane, and degassed by sonication or vacuum to prevent air bubbles in the purification column, which could affect the separation efficiency.
• Before use, the column must be fully equilibrated to room temperature (18-25℃); temperatures too low or too high can impact exosome isolation efficiency.
• The column must be bubble-free. Thoroughly check before and after use to ensure optimal performance.
• The column can be reused, but multiple uses may affect purification quality. It is recommended not to exceed five uses.
• For NGS or other omics analyses, use a new purification column for each sample to prevent cross-contamination.

Advantages:

• High purity and recovery rate of isolated exosomes.
• Simple operation, no specialized purification equipment required.
• The isolated exosomes are free from protein contamination, making them suitable for identification, detection, and functional research.

III. Spin Column Exosome Isolation Kit

	Catalog No.: P-CA-504  Kit Components:  - Spin Column 01  - Collection Tubes (2 mL)  - Solution A

Principle Overview:

The spin column is pre-packed with specific media that binds impurities but not exosomes, allowing for efficient separation through centrifugal force. This method enables quick isolation of exosomes while effectively removing free labeling dyes.

Workflow:

1. Sample Pretreatment: Perform differential centrifugation (300×g, 5 min → 2000×g, 10 min → 14000×g, 30 min) to gradually remove cells, debris, and large particles.

2. Column Preparation: Discard the protective liquid from the spin column and wash with Solution A.

3. Exosome Isolation: Add the pretreated sample to the column, centrifuge, and collect the eluate containing isolated exosomes.

Precautions:

• The maximum loading volume for a single spin column is 500 μL. If the sample volume exceeds 500 μL, it is recommended to use a 50 kDa ultrafiltration tube to concentrate the sample to 0.5 mL, ensuring the concentration volume does not exceed 20 times.
• For high-viscosity samples such as plasma, serum, pleural effusion, and ascites, it is recommended to dilute with an equal volume of deionized water before loading onto the spin column.
• For samples with high impurity content, such as plasma, serum, pleural effusion, and milk, dilute with PBS at a 1:10 ratio, then concentrate tenfold using a 50 kDa ultrafiltration tube. Add tenfold the remaining liquid volume with PBS, concentrate again tenfold, and finally use the spin column for exosome isolation.
• When performing centrifugal separation, maintain the centrifugal force at 300-500×g. For highly viscous samples, the centrifugal force can be appropriately increased to ensure proper separation.

Advantages:

• Simple operation with high purity and high enrichment.
• No need for magnetic racks, ultracentrifuges, or other specialized equipment.
• Efficiently removes free labeling dyes from isolated exosomes.

IV. Exosome Isolation Kit (Precipitation)

	Catalog No.: P-CA-505  Kit Components:  - Exosome Precipitation Solution  - Solution A

Principle Overview:

By adding a specific reagent (Exosome Precipitation Solution), exosomes in the sample rapidly aggregate and precipitate. Exosomes are then collected through centrifugation, providing a quick and efficient method for isolation.

Workflow:

1. Sample Pretreatment: Perform differential centrifugation (300×g, 5 min → 2000×g, 10 min → 14000×g, 30 min) to gradually remove cells, debris, and large particles.

2. Exosome Precipitation:

a) For serum/plasma samples, mix the sample, Solution A, and Exosome Precipitation Solution in a 2:2:1 volume ratio.

b) For cell culture supernatants or urine samples, mix the sample and Exosome Precipitation Solution in a 4:1 volume ratio.

3. Incubate at room temperature for 30 minutes or overnight at 4°C.

4. Exosome Recovery: Centrifuge at high speed, discard the supernatant, resuspend the pellet with Solution A, and centrifuge again to collect the supernatant, which contains the isolated exosomes.

Precautions:

• Pay attention to the volume ratio of the sample and reagents; follow the recommended ratio strictly.
• For large volumes of cell culture supernatants or urine samples, concentration before precipitation is recommended.
• Handle samples gently during pipetting to avoid disturbing the pellet, which may affect the yield.
• If time allows, incubating overnight at 4°C is recommended for optimal recovery rates.

Advantages:

• Suitable for a wide range of sample types with high recovery rates and reproducibility.
• Simple and rapid operation with no need for specialized equipment.
• High throughput, capable of processing multiple samples simultaneously.

Whether you are working with rare micro samples or need large-scale, high-throughput processing, Pricella® Exosome Isolation Kits guarantee high-purity, high-yield, and user-friendly solutions. Pricella® is committed to continuously optimizing product performance to support exosome research and enhance applications in disease diagnostics.