Product Overview
The APD-07 Chicken scFv Phage Display Library Construction Kit is designed for the construction of chicken IgY-derived single-chain variable fragment (scFv) antibody phage display libraries. The kit includes chicken antibody variable-region amplification and assembly primer mixes together with the pAPD-ck-scFv phagemid vector.
Chicken VH and VL antibody repertoires can be amplified from cDNA prepared from peripheral blood lymphocytes or lymphoid tissues of immunized or non-immunized donors. The amplified repertoires are assembled into VH-(G4S)3-linker-VL scFv constructs by overlap-extension PCR.
The vector supports two restriction-enzyme cloning methods: Method A (SacI/SpeI) and Method B (SfiI).
Chicken scFv Library Construction Workflow
- Variable-region amplification: Amplify chicken VH and VL repertoires using the supplied amplification primer mixes.
- scFv assembly: Assemble the VH and VL repertoires into chicken scFv constructs using overlap-extension PCR and the supplied assembly primer mix.
- Restriction digestion: Digest the assembled scFv repertoires and pAPD-ck-scFv vector using either SacI/SpeI (Method A) or SfiI (Method B).
- Ligation and transformation: Purify the digested DNA fragments, ligate the chicken scFv inserts into the corresponding phagemid vector, and transform competent bacterial cells.
- Phage display: Prepare the chicken scFv phage display library for downstream biopanning, antibody selection, and screening.
Key Features
- Chicken antibody library construction: Designed for chicken IgY-derived VH and VL repertoires.
- Complete primer system: Includes chicken variable-region amplification and scFv assembly primer mixes.
- Flexible cloning options: Supports SacI/SpeI and SfiI restriction-enzyme cloning.
- Phagemid vector: Includes the 5,349 bp pAPD-ck-scFv vector for chicken scFv phage display library construction.
- Downstream applications: Suitable for antibody phage display, biopanning, and antibody candidate screening.
Specifications of Antibody Phage Display Vector
Specifications of Antibody Phage Display Vector| Antibiotic Resistance | Ampicillin (AmpR) |
| Constitutive or Inducible System | Inducible for downstream expression |
| Delivery Type | Transformation |
| Product Type | Phage display vector or Bacterial Expression vector |
| Cloning Method | Either [Both 5’end and 3’end SfiI sites] or [5’end SacI and 3’ end SpeI sites] |
Contents & Storage
The kit contains six vials for chicken scFv phage display library construction.
Content of chicken scFv phage display library construction Kit| Primer for amplification of chicken variable domain of heavy chain (VH) and light chain (VL) | ||
| Vial 1 | 100 µl, 10 µM | Primers (normalized forward and reverse primer mix) for VH repertoires amplification |
| Vial 2 | 100 µl, 10 µM | Primers (normalized forward and reverse primer mix) for VL repertoires amplification |
| Primer Set for Cloning/assembly chicken scFv (Format: VH-(G4S)3 linker-VL) | ||
| Vial 3 | 100 µl, 10 µM | Primers (Forward primer and reverse mix) for chicken scFv repertoires assembly |
| Sequencing Primer set | ||
| Vial 4 | 100 µl, 10 µM | M13 Reverse primer for scFv forward sequencing |
| Vial 5 | 100 µl, 10 µM | pIII Reverse primer for scFv reverse sequencing |
| pAPD-ck-scFv cloning vector for phage display chicken scFv library construction | ||
| Vial 6 | 10.0 µg in Tris-EDTA buffer | |
Storage: Store all kit components at -20°C. The primer sets and vector are guaranteed stable for 12 months when properly stored.
Vector Usage
- Construction of chicken scFv antibody phage display libraries.
- Cloning of chicken scFv repertoires into the pAPD-ck-scFv phagemid vector.
- Antibody phage display selection, biopanning, and downstream antibody screening.
