PBMC Isolation and Processing for Research and Clinical Trials
Accelevir is a CLIA-certified lab specializing in high-quality PBMC isolation from a variety of sources. Our end-to-end services—from isolation and enrichment to cryopreservation and downstream analysis—support researchers and drug developers advancing immunological, oncological, and gene therapy programs.
Our Differentiators:
- CLIA-certified, GCLP-aligned workflows
- Multiple isolation techniques: Ficoll, SepMate™, CPT, RoboSep™, EasySep™
- Expertise in leukopak, whole blood, and buffy coat processing
- Custom protocols for cell subset enrichment and dry pellet prep
- Central lab logistics, rapid turnaround, and global support
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Introduction to PBMCs
Peripheral Blood Mononuclear Cells (PBMCs) include immune subsets such as:
- Lymphocytes: T cells, B cells, NK cells
- Monocytes
- Dendritic cells
Typical Cell Frequencies
PBMCs make up ~1% of whole blood. Subsets vary by individual and disease state.
Why PBMCs Matter
- Immunoassays: ELISpot, cytokine release
- Gene & Cell Therapy: AAV/LV response and persistence
- Clinical Biomarker Discovery
- Toxicology Studies: Immune response to compounds
- Rare Disease Research
PBMC Isolation Methods Offered
Ficoll-Paque Density Gradient
- Standard method
- 45–60 min per sample
- Low cost, consistent viability
SepMate™ Tubes
- Built-in barrier improves layering
- Speeds up DGM: ~15 mins/sample
- Reduces variability
CPT Tubes
- Pre-filled with anticoagulant and gel
- Enables field or clinical site centrifugation
- Maintains integrity during transport
EasySep™ Magnetic Cell Separation
- RBC/granulocyte depletion
- No centrifuge needed
- Preserves subset function
RoboSep™ Automation
- Fully automated for PBMC prep
- Ideal for large-volume, standardized studies
PBMC Isolation Protocols & Best Practices
Core Workflow
- Sample log-in and ID verification
- DGM or magnetic separation
- RBC/granulocyte depletion (if needed)
- PBMC washing and pelleting
- Cell viability check (trypan blue or AO/PI)
- Cryopreservation or immediate analysis
Equipment
- Class II BSC
- Swing-bucket centrifuges
- Cell counters (manual + automated)
- Vapor-phase LN₂ tanks
- Label printers, cryovial systems
Lab Requirements
- BSL-2 certified lab
- Trained personnel with SOPs
- 21 CFR Part 11 sample tracking
Cell Counting and Viability Metrics
Manual Counting (Trypan Blue)
- Hemocytometer-based
- Detects viability via membrane integrity
Automated Counting (AO/PI)
- Higher throughput
- Enhanced standardization
Routine results: >90% viability across sample types
Cryopreservation & Thawing Protocols
Cryopreservation Process
- Controlled freezing: −1°C/min ramp
- DMSO/FBS or CryoStor® CS10 media
- Final concentration: 5–10×10⁶ cells/mL
- Storage at −80°C then liquid nitrogen
Thawing Best Practices
- 37°C rapid thaw
- Gradual dilution to reduce osmotic shock
- Centrifugation and wash to remove DMSO
PBMC Sources Supported
- Whole blood: EDTA, Li-Heparin, CPT
- Buffy coat: separated from whole blood
- Leukopaks: large-scale apheresis products
- LRS cones: leukocyte-depleted filter products
- Cord blood / Bone marrow: upon request
Immune Cell Subset Enrichment
Magnetic Enrichment
- CD3/CD4/CD8/CD14/CD19/CD56/CD34
- Untouched or positively selected options
Plastic Adherence
- Monocyte enrichment using flask adhesion
- Effective alternative for certain cytokine studies
Comparative PBMC Isolation Metrics
| Method | Time | Viability | Contamination | Volume Scale |
| Ficoll | ~60 min | High | Moderate (platelets, RBCs) | Moderate |
| SepMate™ | ~15 min | High | Low | Moderate |
| CPT | ~30 min | High | Variable | High (clinical sites) |
| EasySep™ | ~20 min | High | Lowest | Small to mid |
| RoboSep™ | ~25 min | High | Lowest | High-volume |
Troubleshooting and Optimization
- Delayed Processing: Use CPT or cryostabilize within 6–8 hours
- Granulocyte Contamination: Minimize sample age; use EasySep™
- Platelet Contamination: Double wash or deplete magnetically
- Temperature Issues: Maintain 18–25°C before processing
- Anticoagulant Mismatch: Match protocol to tube type
Downstream Applications
- Flow Cytometry: Immune phenotyping
- Functional Assays: ELISpot, cytokine profiling
- Transcriptomics: scRNA-seq, qPCR
- Drug Screening: Cell-based assays
- Gene Therapy: AAV/LV transduction analysis
PBMC Products & Custom Services
- Fresh/frozen PBMCs with donor spec
- Cryopreserved lots for longitudinal studies
- Isolation from leukopaks, whole blood, cones
- Donor screening (HLA, CMV, BMI, age, etc.)
- Custom protocols for preclinical and clinical trials
Related Services (Internal Links)
- Flow Cytometry Services
- Molecular & Genomic Assays
- Custom Assay Development
- Immunogenicity Testing
- Gene Therapy Analytics
- Start a Project
FAQs
Do you support automated PBMC isolation?
Yes, with RoboSep™ for high-throughput workflows.
Can I source cryopreserved PBMCs with donor spec?
Yes, including HLA, CMV status, BMI, and more.
Which anticoagulants are compatible?
We accept EDTA, ACD, Heparin, CPT. Reach out for compatibility confirmation.
What is your viability guarantee?
90% post-thaw for validated workflows.
Are custom enrichment protocols available?
Yes. We provide magnetic and adherence-based methods.
Request a Quote or Partner With Us
Ready to start your PBMC isolation project? Partner with Accelevir for trusted, high-quality, CLIA-certified services.