At a glance: Characteristics of the AbNano™ VHH Naïve Library
| 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 to 1 x 1011 PFU/mL and aliquoted as 1 mL tubes frozen and stored at -80 °C. The library is delivered as 1 mL aliquots of 1 x 1011 PFU/mL in PBS + 20% glycerol and can be used to pan for up to 40 targets.
