Asymmetrical Flow Field-Flow Fractionation (AF4) Services
ZentriForce Pharma provides Asymmetrical Flow Field-Flow Fractionation (AF4) services as part of its orthogonal portfolio for biopharmaceutical characterisation.
AF4 Services for Advanced Biotherapeutic Modalities Characterisation Across a Broad Size Range
AF4 separates biologics particles directly in solution, covering a broad measurement range from 1 nm to 1 μm — ideal for molecules and complexes that extend beyond the scope of SEC-MALS or SV-AUC alone.
By integrating AF4 with detectors such as Multi-Angle Light Scattering (MALS) and Refractive Index (RI), the company delivers precise data on particle size distributions, soluble aggregates, oligomeric species, and structural integrity. Regulatory agencies, including the FDA and EMA, increasingly value AF4 as a complementary method to SEC, making it an essential tool for development and biosimilarity studies.
Why Choose AF4 Services at ZentriForce Pharma?
- Broad measurement scope: Covers 1 nm – 1 μm, addressing peptides, proteins, nucleic acids, nanoparticles, LNPs, viruses, and liposomes.
- Orthogonal approach: Complements SEC-MALS and SV-AUC for a complete picture of sample purity and stability.
- Advanced instrumentation: Equipped with the Wyatt Eclipse AF4 system integrated with Agilent 1260 Infinity II HPLC, Wyatt DAWN Heleos II MALS, and Optilab T-rEX RI detector for high sensitivity and reproducibility.
- Innovative data analysis: ASTRA software for standard AF4-MALS data analysis and interpretation.
- Efficient workflows: Cost-effective method development supported by extensive expertise in biosimilarity and gene therapy projects.
- Regulatory alignment: Increasingly recognised by the FDA and EMA as an orthogonal tool to support SEC findings.
Technical Specifications
| Parameter | Specification |
| Instrument | Wyatt Eclipse AF4 system with a Frit-Inlet channel |
| Integrated detectors | Agilent 1260 Infinity II HPLC, Wyatt DAWN Heleos II MALS, Wyatt Optilab T-rEX RI |
| Size range | 1 nm – 1 μm |
| Applications | Particle size distribution, aggregate detection, oligomer resolution, species characterisation, protein conjugate and antibody-drug conjugate (ADC) analysis |
| Sample fractionation | Equipped with a bioinert fraction collector for offline analysis of separated species (e.g., bioassays, mass spectrometry) |
| Data analysis | ASTRA software for AF4-MALS data analysis and evaluation |
Applications of AF4
Gene Therapy Development
AF4 enables precise size-based separation across DNA/RNA complexes, adeno-associated viruses (AAV), and larger viruses. This broad coverage makes it a key method for evaluating critical quality attributes of gene therapy products.
Biosimilarity Studies
AF4 provides orthogonal confirmation of SEC-MALS findings, offering high-resolution analysis of soluble aggregates, oligomers, and species distributions. ZentriForce Pharma has applied AF4 in multiple biosimilarity projects, including peptides, antibodies, and oligonucleotides, helping clients generate robust data packages for regulatory submissions.
Biopharmaceutical Development
With its wide dynamic range, Asymmetrical Flow Field-Flow Fractionation (AF4) is highly effective for process optimisation and troubleshooting. It provides detailed information on aggregate quantification, particle size distributions, and structural assessments, supporting both early development and quality control.
Quality and Expertise You Can Trust
- BSL-1 and BSL-2 facilities for handling a diverse range of biopharmaceutical materials.
- Comprehensive quality system aligned with WHO guidelines, supported by procedural, equipment, and analytical SOPs.
- Dedicated quality manager overseeing compliance, staff training, and equipment qualification.
- Audit-ready environment with transparent processes and full data traceability.
Partner with ZentriForce Pharma
By combining AF4 with complementary methods such as SEC-MALS, SV-AUC, and μ-DSC, ZentriForce Pharma delivers a broad suite of advanced analytical solutions for biopharmaceutical development.
📩 Contact ZentriForce today to learn how AF4 can support your biosimilarity, gene therapy, or biopharmaceutical project. Please email ZentriForce’s experts at info@zentriforce.com or complete the Quick Contact Form.
