Product Overview
Fusion BioLabs offers antibody phage display library construction kits featuring specialized PCR primer sets and phagemid vectors for generating antibody libraries. These components support the construction of customized antibody phage display libraries for antibody discovery, screening, and affinity maturation research.
pAPD-h-scFv is a phagemid vector designed for constructing human single-chain variable fragment (scFv) antibody libraries. The general library construction workflow includes the following steps:
- Amplify antibody variable-region genes from cDNA prepared from RNA isolated from peripheral blood lymphocytes (PBL) or lymphoid tissues of non-immunized or immunized donors. PCR primers corresponding to known VH, Vκ, and Vλ gene sequences are used.
- Combine VH and VL repertoires to generate scFv constructs using a two-fragment PCR assembly procedure.
- Digest the pAPD-h-scFv vector and assembled scFv repertoires with the restriction enzyme SfiI.
- Ligate the digested and purified scFv fragments into the corresponding SfiI-digested and purified pAPD-h-scFv vector.
Key Features
- Efficient scFv display: Designed to support expression and phage display of scFv antibody fragments for screening and selection against target molecules.
- Flexibility and versatility: The pAPD-h-scFv vector supports both antibody phage display library construction and downstream antibody fragment expression, without requiring subcloning into a separate expression vector for compatible applications.
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 | Both 5’end and 3’end SfiI sites |
Contents & Storage
Content of human scFv phage display library construction Kit| scFv Construction Primer Set | ||
| Vial 1 | 100 µl, 10 µM | Primer mix (F+R) for Vk repertoires amplification |
| Vial 2 | 100 µl, 10 µM | Primer mix (F+R) for Vλ repertoires amplification |
| Vial 3 | 100 µl, 10 µM | Primer mix (F+R) for VH repertoires amplification |
| Vial 4 | 100 µl, 10 µM | Assembly primers (F+R) for scFv repertoires construction |
| pAPD-h-scFv cloning vectors for human scFv library construction | ||
| Vial 5 | 10.0 µg in Tris-EDTA buffer | |
| Sequencing Primer set | ||
| Via 6 | 100 µl, 10 µM | M13 Reverse primer for scFv forward sequencing |
| Via 7 | 100 µl, 10 µM | pIII Reverse primer for scFv reverse sequencing |
- Storage: Store at -20°C. Vectors are guaranteed stable for 12 months when properly stored.
Vector Usage
- Cloning scFv inserts into the pAPD-h-scFv vector for antibody phage display library construction.
- Cloning selected scFv candidates into the pAPD-h-scFv vector for downstream antibody fragment expression.
