- Format: VHH with HA Tag and 6xHis Tag (pelB leader sequence-VHH-HA tag-6xHis tag)
- Framework: ADE99145
- Mutated: CDR1, CDR2 and CDR3 (Saturation mutagenesis)
- Diversity: 8.9×1010
- The diversity and in-frame of the library was checked by NGS.
The library was constructed by saturation mutagenesis of all 3 CDRs and their CDR combination. The main library consists of 5 sub-libraries with the following CDR composition and saturation mutated CDR combination.
Composition/distribution of CDRs of five (5) VHH sub-libraries
| Sub-libraries | CDR1 (7 amnion acids) | CDR2 (6 amino acids) | CDR3 (17 or 23 amino acids) |
|---|---|---|---|
| 1 | Saturation Mutagenesis | Saturation Mutagenesis | M17 Saturation Mutagenesis |
| 2 | Saturation Mutagenesis | Saturation Mutagenesis | M23 Saturation Mutagenesis |
| 3 | No Mutagenesis | No Mutagenesis | M17 Saturation Mutagenesis |
| 4 | No Mutagenesis | No Mutagenesis | M23 Saturation Mutagenesis |
| 5 | No Mutagenesis | Saturation Mutagenesis | M23 Saturation Mutagenesis |
- Kit contents
The following components are included in the Kit.
Ready-to-panning Synthetic VHH Library Kit Content
| Component | Quantity | Composition |
|---|---|---|
| Synthetic VHH library (ready-to-panning phage; 8.0×1012pfu/ml) | 1.0 ml | 1×PBS with 25% glycerol |
| M13KO7 Helper Phage (2.0×1012 pfu/ml) | 0.5 ml | 1×PBS with 25% glycerol |
| Chemically Competent TG1 E. coli (2.5x108cfu/ml) | 0.5 ml | 2xYT with 25% Glycerol |
| M13 Reverse Primer (1.6 µM) | 0.2 ml | 1×TE Buffer |
| pIII Reverse Primer (1.6 µM) | 0.2 ml | 1×TE Buffer |
