Product Overview
Fusion BioLabs offers a range of library primer sets and phagemid vector combination 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-m-Fab is the phagemid vector for construction of a fragment antigen-binding (Fab) library for mouse antibodies. Here are the key steps involved in constructing such a library:
- Amplify V genes from cDNA reverse transcript 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 repertoires and CH1 fragment, and VL repertoires and CL fragment to create VH-CH1 and Vk,λ-Ck,λ constructs respectively, using a simple two-fragment PCR assembly procedure.
- Restriction enzyme digestion pAPD-m-Fab vector and Vk,λ-Ck,λ fragments with SacI/XbaI, or pAPD-m-Fab vector and VH-CH1 fragments with XhoI/SpeI.
- Ligation of digested and purified fragment into corresponding restriction enzymes digested and purified pAPD-m-Fab vector to make either Light chain sub-library or heavy chain sub-library.
- Restriction enzyme digestion light chain sub-library and VH-CH1 fragment with XhoI/SpeI or heavy chain sub-library and Vk,λ-Ck,λ fragment with SacI/XbaI, to make mouse Fab libraries.
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 subcloning into 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 | 5’-SacI and 3’-SpeI |
Contents & Storage
Content of mouse Fab phage display library construction Kit| Primer Set | ||
| Vial 1 | 100 µl, 10 µM | Forward Primer mix (14 oligos) for Vk,λ-Ck,λ fragment amplification |
| Vial 2 | 100 µl, 10 µM | Reverse Primer mix (2 oligos) for Vk,λ-Ck,λ fragment amplification |
| Vial 3 | 100 µl, 10 µM | Forward Primer mix (11 oligos) for VH-CH1 fragment amplification |
| Vial 4 | 100 µl, 10 µM | Reverse Primer mix (3 oligos) for VH-CH1 fragment amplification |
| Sequencing Primer set | ||
| Vial 5 | 100 µl, 10 µM | M13 Reverse primer for scFv forward sequencing |
| Vial 6 | 100 µl, 10 µM | pIII Reverse primer for scFv reverse sequencing |
| pAPD-m-Fab vector for mouse Fab phage display library construction | ||
| Vial 7 | 10.0 µg in Tris-EDTA buffer | |
- Store at -20°; Vectors are guaranteed stable for 12 months when properly stored.
Vector usage
- Cloning Fab inserts into pAPD-m-Fab vector for phage display library construction.
- Cloning Fab candidate into pAPD-m-Fab vector for antibody fragment expression.
