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AbNano™ VHH Libraries

AbNano™ VHH Naïve Library from Fortis Life Sciences® is a fully natural, single domain naïve library with source material from camelids and the antibodies displayed on monovalent phage. It is intended for research purposes only to aid in the discovery of antibodies intended for use as therapeutics, diagnostics, and research tools.

This fully natural library has higher diversity in framework regions than synthetic libraries. Naïve libraries have the potential to contain binders to a wide variety of targets, and the distribution of CDRH3 length in our library is similar to human length distributions, which may improve the potential for therapeutic application of these VHH. We have already discovered nanomolar binders to key therapeutic targets such as PD-L1, EGFR, and INSR-β.

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Single-domain antibodies or VHH domains

At a glance: Characteristics of the AbNano™ VHH Naïve Library

Characteristic

Details

Affinity of typical hits on protein target

10 nM – 1 μM

Downstream applications

Multispecific formats
Cell recruitment

Number of unique targets

Many

Number of animals used

103

PBMC source material

>1E10

Timeline to results

As little as 1 month

AbNano™ VHH Naïve Library Specifications

The library was constructed from 103 naïve animals: 77 llamas and 26 alpacas. A total of 1.51 x 1010 PBMC cells were collected and used for library construction.

Libraries were constructed following Abcore's internal workflows for RNA isolation, cDNA synthesis, and PCR amplification using species-specific primers. Transformants were analyzed initially by Sanger sequencing. Sanger analysis demonstrated 89% of VHH sequences in-frame. The library was subsequently analyzed by next generation sequencing methodology. Reads were clustered and analyzed for diversity and confirmed to meet our QC standards.

The total number of transformants for the library was 1.12 x 1010 CFU. Library bacterial stocks were grown to log phase, infected by M13KO7, PEG precipitated, and the pooled phage was titered. Phage were normalized in PBS + 20% glycerol and aliquoted as 1 mL tubes frozen and stored at -80 °C. The library is delivered as 1 mL aliquots.

Nanomolar VHH binders of PD-L1 and other therapeutic targets from the AbNano™ VHH Naive Library

Discovering novel VHH domains can take place by multiple workflows, including using B-cell sorting and using display libraries. In this poster, we present the construction and validation of the AbNano™ VHH Naive Library, a large naïve library from llamas and alpacas.


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