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-Fab is a phagemid vector designed for constructing human antigen-binding fragment (Fab) 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.
- Amplify the CH1, Cκ, and Cλ constant-region fragments using pAPD-h-Fab as the PCR template.
- Combine VH repertoires with CH1, Vκ repertoires with Cκ, and Vλ repertoires with Cλ to generate VH–CH1, Vκ–Cκ, and Vλ–Cλ constructs, respectively, using a two-fragment PCR assembly procedure.
- Perform overlap assembly of Vκ–Cκ with VH–CH1, or Vλ–Cλ with VH–CH1, to generate the corresponding human Fab repertoires.
- Digest the pAPD-h-Fab vector and assembled Fab repertoires with the restriction enzyme SfiI.
- Ligate the digested and purified Fab repertoires into the corresponding SfiI-digested and purified pAPD-h-Fab vector to construct human Fab phage display libraries.
Key Features
- Efficient Fab display: Designed to support expression and phage display of Fab antibody fragments for screening and selection against target molecules.
- Flexibility and versatility: The pAPD-h-Fab 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 | Either [Both 5’end and 3’end SfiI sites] or [5’end SacI and 3’ end SpeI sites] |
Contents & Storage
Content of human Fab phage display library construction Kit| Primer Set | ||
| Vial 1 | 100 µl, 10 µM | Primer mix (F+R) for VH repertoires amplification |
| Vial 2 | 100 µl, 10 µM | Primer mix (F+R) for Vk repertoires amplification |
| Vial 3 | 100 µl, 10 µM | Primer mix (F+R) for Vλ repertoires amplification |
| Vial 4 | 100 µl, 10 µM | Primer mix (F+R) for CH1 fragment amplification |
| Vial 5 | 100 µl, 10 µM | Primer mix (F+R) for Ck fragment amplification |
| Vial 6 | 100 µl, 10 µM | Primer mix (F+R) for Cλ fragment amplification |
| Vial 7 | 100 µl, 10 µM | Assembly primers for VH-CH1 repertoires construction |
| Vial 8 | 100 µl, 10 µM | Assembly primers for Vk-Ck repertoires construction |
| Vial 9 | 100 µl, 10 µM | Assembly primers for Vλ-Cλ repertoires construction |
| Vial 10 | 100 µl, 10 µM | Assembly primers for Fab repertoires construction |
| Vial 11 | 100 µl, 10 µM | M13 rev Forward sequencing primer |
| Vial 12 | 100 µl, 10 µM | pIII rev Reverse sequencing primer |
| pAPD-h-Fab cloning vectors for human Fab library construction and templates for CH1, Ck and Cλ fragment amplification | ||
| Vial 13 | 10.0 µg in Tris-EDTA buffer | |
- Storage: Store at -20°C. Vectors are guaranteed stable for 12 months when properly stored.
Vector Usage
- Cloning Fab inserts into the pAPD-h-Fab vector for antibody phage display library construction.
- Cloning selected Fab candidates into the pAPD-h-Fab vector for downstream antibody fragment expression.
