Product Overview
Fusion BioLabs offers a range of library primer sets and phagemid vector combinations for antibody phage display and peptide phage display construction. With customizable features and robust performance, our primer sets and phagemid vectors are designed for facilitating phage display library generation as fast as within one week.
pAPD-h-scFv is the phagemid vector for construction of a single-chain variable fragment (scFv) library for human antibodies. Here are the key steps involved in constructing such a library:
- Amplify V genes from cDNA reverse transcribed from RNA isolated from peripheral blood lymphocytes (PBL) or lymphoid tissue of non-immunized or immunized donors using PCR primers corresponding to known VH, Vκ, and Vλ gene sequences.
- Combine VH and VL repertoires to create the scFv construct using a simple two-fragment PCR assembly procedure.
- Restriction enzyme digestion of the pAPD-h-scFv vector and scFv repertoires with SfiI.
- Ligation of digested and purified scFv fragment into corresponding restriction enzyme(s) digested and purified pAPD-h-scFv
Key Features
High expression efficiency: Engineered for efficient expression and display of antibody fragment scFv on the surface, allowing for easy screening and selection of target molecules.
Flexibility and versatility: One vector for both antibody library construction and downstream antibody fragment expression. No need to subclone into an expression vector for downstream application.
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 | Restriction Enzyme (5’-SacI and 3’-SpeI) |
Contents & Storage
Content of human scFv phage display library construction Kit| scFv Construction Primer Set | ||
| Vial 1 | 200 µl, 10 µM | Primer mix (F+R) for Vk repertoires amplification |
| Vial 2 | 200 µl, 10 µM | Primer mix (F+R) for Vλ repertoires amplification |
| Vial 3 | 200 µl, 10 µM | Primer mix (F+R) for VH repertoires amplification |
| Vial 4 | 200 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 |
- Store at -20°. Vectors are guaranteed stable for 12 months when properly stored.
Vector usage
- Cloning scFv inserts into the pAPD-h-scFv vector for phage display library construction.
- Cloning scFv candidate into pAPD-h-scFv vector for antibody fragment expression.
