Can Nanobodies Innovate Large-Scale Vaccine Purification?
Introduction
Researchers have developed NbHB1, a camelid-derived nanobody, to purify recombinant Hepatitis B surface antigen virus-like particles (rHBsAg-VLPs) with large scale performance. This approach combines high specificity, excellent reusability, and scalability — all essential for modern vaccine production.
Research Highlights
How It Works
- Expression: NbHB1 was produced in E. coli as inclusion bodies.
- Refolding: Recovered through an efficient two-step refolding strategy.
- Immobilization: Fused with a SpyTag and covalently bound to a SpyCatcher-derived support, creating a nanobody-liganded affinity resin.
Performance Highlights
Purification Efficiency
- Static Binding Capacity: 2.03 mg/mL
- Dynamic Binding Capacity: 1.43 mg/mL at 10% breakthrough
- Purity: 96% in a single step
- Recovery: 81% yield from clarified CHO cell culture fluid
Scalability
- Performance was identical when scaling up from 5 mL to 500 mL columns.
Reusability
- 25 purification cycles without loss of capacity
- Ligand Leakage: Below 1.7 ng/mL resin across all cycles
Chemical Stability
- Stable with 0.1 M citric acid, 20 mM NaOH, and 2 M guanidine chloride.
Why It Matters
Traditional antibody-based affinity resins can be costly, less stable, and prone to leaching. Nanobodies like NbHB1 solve these issues by:
- Offering higher stability under harsh cleaning conditions.
- Maintaining binding performance over multiple cycles.
- Enabling cost-effective large-scale purification.
Future Applications
While NbHB1 is designed for Hepatitis B vaccine production, the platform can be adapted for:
- Other virus-like particle vaccines (HPV, SARS-CoV-2 VLPs).
- Recombinant protein biologics.
- Gene therapy vectors where high-purity, high-yield purification is critical.
Conclusion
The NbHB1 nanobody-liganded affinity resin delivers a 96% purity, 81% recovery, and 25-cycle durability — setting a new benchmark for industrial vaccine purification. Its combination of efficiency, scalability, and robustness could accelerate vaccine manufacturing worldwide.
Read the full study here: https://www.sciencedirect.com/science/article/pii/S0021967325001335