Efficient discovery of high-affinity, stable, and easily producible VHHs (nanobodies) remains a major challenge in biopharmaceutical research. AlpVHHs leverages advanced phage display technology to empower precise VHH antibody discovery, enabling faster development of therapeutic and diagnostic antibodies.
Through a mature CRO workflow, we transform immune libraries into high-affinity nanobody candidates, offering reliable, traceable technical support. Our expertise streamlines the target-to-candidate process, significantly accelerating drug discovery timelines.
Phage display is a molecular technique where peptides, antibodies, or protein fragments are fused to phage coat proteins (pIII or pVIII). The displayed molecules retain their biological activity, allowing high-throughput screening against target antigens.
The selection process, known as biopanning, involves:
Incubation – Phage library is exposed to the target antigen to form stable complexes.
Washing – Weak or non-specific binders are removed, enriching high-affinity candidates.
Elution – Specific phages are recovered for further amplification.
Amplification – E. coli infects eluted phages, producing a progeny library for subsequent rounds.
After 2–3 rounds of this adsorption–elution–amplification cycle, high-affinity clones dominate, ready for downstream validation.
First reported by George P. Smith in 1985 and later advanced by Sir Gregory Winter, phage display technology earned them the 2018 Nobel Prize in Chemistry, underscoring its critical role in modern antibody engineering and therapeutic discovery.
High-Throughput Screening: Billions of variants displayed simultaneously, accelerating candidate identification.
Stability: Phage libraries can be stored long-term without losing diversity.
Flexibility: Suitable for a wide range of molecules, including full-length proteins, scFvs, VHHs, and peptides.
5' RACE-based Optimization: Refined primers maximize sequence coverage and reduce amplification bias.
Zero-Background Technology: Ensures 100% library insertion rate.
Synergistic Platform: Combines homologous recombination with ultra-competent cells for library titers of 10⁹.
High-Efficiency Transformation: Proprietary electro-transformation protocols achieve 10⁹ diversity in a single reaction.
Adaptive Screening Strategies: Flexible methods ensure a >90% project success rate.
Proven Track Record: Over 800 completed VHH development projects demonstrate deep expertise.
AlpVHHs ensures high-quality phage libraries through rigorous optimization and validation, supporting efficient VHH antibody discovery:
Primer Optimization: Proprietary primers provide broad coverage of HcAb germline sequences, maximizing diversity.
Zero-Background Cloning: Achieves 100% VHH insertion efficiency with background-free cloning, eliminating empty clones.
Large Library Size: Guaranteed library capacity exceeding 1×10⁹ CFUs, enabling extensive screening.
Clonal Uniqueness: 100% uniqueness confirmed by Sanger sequencing of 48 random clones, ensuring diverse candidate selection.
High ORF Accuracy: Validated clones achieve >90% correctness rate in open reading frames, minimizing screening bias.
These rigorous controls result in diverse, high-quality alpaca VHH phage display libraries that significantly improve downstream screening success rates.
Prokaryotic Expression Bias – Antibodies of eukaryotic origin may face codon bias or toxicity in E. coli.
Library Bias – Empty clones or incorrect ORF frames can reduce library quality.
Large Molecule Constraints – Phage display struggles with very large proteins.
Resource Intensive – High-quality library construction requires specialized skills and time.
Customized immunization strategies and primer optimization minimize expression bias.
Zero-Background Technology eliminates interference from empty clones.
End-to-end guidance ensures high success rates in library construction and screening, even for complex targets.
AlpVHHs has successfully applied phage display to multi-pass transmembrane proteins and tumor-associated antigens, generating high-affinity, high-specificity nanobody candidates.
Target examples successfully addressed include:
| Multi-pass transmembrane proteins: | CCR1, CCR8, CXCR3, GPRC5D, TSHR, CLDN18.2, CLDN4 |
|---|---|
| Tumor-associated antigens: | BCMA, CD19, CD20, PSMA, CD33, CD38, EpCAM, and others |
Target: CLDN18.2, critical for gastric and pancreatic tumors.
Immunization: Alpacas achieved serum titers of 1:64K.
Library Diversity: 2.17 × 10⁹.
Screening: Three rounds of cell-based panning; 175 monoclonal clones yielded 56 target-specific clones.
Outcome: 16 unique CDR sequences identified; 10 showed high-specificity binding to CLDN18.2 without cross-reactivity to CLDN18.1.
These VHH candidates provide a strong foundation for ADC, CAR-T, or bispecific antibody development.
Target: GPRC5D, relevant for multiple myeloma.
Immunization: Alpacas with GPRC5D-overexpressing cells.
Library Diversity: 2.65 × 10⁹.
Screening: 3 rounds yielded 338 clones; 49 unique CDR sequences with 18 distinct CDR3 sequences.
These candidates support affinity maturation and functional validation.
AlpVHHs is a leading antibody discovery platform, offering comprehensive CRO services:
Extensive Project Experience: 800+ VHH development projects completed.
Robust Resources: Proprietary breeding facilities with 800+ camelids enable large-scale immunization programs.
End-to-End Solutions: From antigen design and immunization to phage display screening and antibody humanization.
Globally Trusted Partner: Supporting 300+ biopharmaceutical companies and research institutions worldwide.
| Service | Delivery | Time |
|---|---|---|
| Library construction | • Bacterial library • Phage display library • Library QC report | 4 weeks |
| Library screening | • Sequences of binders • Project report • Library QC report | 4 weeks |
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