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-m-Fab is a phagemid vector designed for constructing mouse fragment antigen-binding (Fab) antibody libraries. The general library construction workflow includes the following steps:
- Amplify mouse VH, Vκ, and Vλ repertoires from cDNA prepared from RNA isolated from mouse lymphoid tissues or other appropriate sources.
- Amplify the CH1, Cκ, and Cλ constant-region fragments using the supplied pAPD-m-Fab vector as the PCR template.
- Combine VH and CH1, Vκ and Cκ, and Vλ and Cλ fragments using two-fragment PCR assembly to generate the corresponding heavy-chain and light-chain repertoires.
- Digest the light-chain repertoires and pAPD-m-Fab vector with SacI/XbaI, or the heavy-chain repertoire and vector with XhoI/SpeI.
- Ligate the purified fragments into the corresponding digested vector to construct a mouse Fab light-chain or heavy-chain sub-library.
- Complete mouse Fab library construction by introducing the complementary heavy-chain repertoire using XhoI/SpeI, or the complementary light-chain repertoire using SacI/XbaI.
Key Features
- Efficient Fab display: Designed to support expression and phage display of mouse Fab antibody fragments for screening and selection against target molecules.
- Flexible library construction: Supports both κ and λ light-chain repertoires and construction through either light-chain or heavy-chain sub-libraries.
- Versatile vector system: The pAPD-m-Fab vector supports antibody phage display library construction and downstream antibody fragment expression.
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 | Restriction Enzymes for (5’-SacI and 3’-XbaI for Vk,λ-Ck,λ repertoires; 5’-XhoI and 3’-SpeI for VH-CH1 repertoire) |
Contents & Storage
Content of mouse Fab phage display library construction Kit| Primer Set | ||
| Vial 1 | 100 µl, 10 µM | Primer mix (F+R) for Vk repertoire amplification |
| Vial 2 | 100 µl, 10 µM | Primer mix (F+R) for Vλ repertoire amplification |
| Vial 3 | 100 µl, 10 µM | Primer mix (F+R) for VH repertoire amplification |
| Vial 4 | 100 µl, 10 µM | Primer mix (F+R) for Ck fragment amplification |
| Vial 5 | 100 µl, 10 µM | Primer mix (F+R) for Cλ fragment amplification |
| Vial 6 | 100 µl, 10 µM | Primer mix (F+R) for CH1 fragment amplification |
| Vial 7 | 100 µl, 10 µM | Assembly Primer mix (F+R) for Vk-Ck repertoire construction |
| Vial 8 | 100 µl, 10 µM | Assembly Primer mix (F+R) for Vλ-Cλ repertoire construction |
| Vial 9 | 100 µl, 10 µM | Assembly Primer mix (F+R) for VH-CCH1 repertoire construction |
| Sequencing Primer set | ||
| Vial 10 | 100 µl, 10 µM | M13 Reverse primer for scFv forward sequencing |
| Vial 11 | 100 µl, 10 µM | pIII Reverse primer for scFv reverse sequencing |
| pAPD-m-Fab vector for mouse Fab phage display library construction | ||
| Vial 12 | 10.0 µg in Tris-EDTA buffer | |
- Storage: Store at -20°C. Primer sets and vectors are guaranteed stable for 12 months when properly stored.
Vector Usage
- Cloning mouse Fab repertoires into the pAPD-m-Fab vector for antibody phage display library construction.
- Cloning selected Fab candidates into the pAPD-m-Fab vector for downstream antibody fragment expression.
