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-h/k-Fab and pAPD-h/λ-Fab are the phagemid vectors for construction of a fragment antigen-binding (Fab) library for human 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.
- Amplify CH1 fragment using either pAPD-h/k-Fab or pAPD-h/λ-Fab as template, Ck fragment using pAPD-h/k-Fab as template and Cλ fragment using pAPD-h/λ-Fab as template.
- Combine VH repertoires and CH1 fragment, and Vk,λ repertoires and Ck,λ fragment to create VH-CH1 and Vk,λ-Ck,λ constructs respectively, using a simple two-fragment PCR assembly procedure.
- Overlap assembly Vk,λ-Ck,λ and VH-CH1 to make Fab
- Restriction enzyme digestion pAPD-h/k-Fab vector or pAPD-h/λ-Fab vector and Fab repertoires with SfiI.
- Ligation of digested and purified repertoires into digested and purified pAPD-h/k-Fab vector or pAPD-h/λ-Fab vector to make human 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 | SfiI |
Contents & Storage
Content of human Fab phage display library construction Kit| Primer Set | ||
| Vial 1 | 200 µl, 10 µM | Forward Primer mix (4 oligos) for Vk fragment amplification |
| Vial 2 | 200 µl, 10 µM | Reverse Primer for Vk fragment amplification |
| Vial 3 | 200 µl, 10 µM | Forward Primer mix (9 oligos) for Vλ fragment amplification |
| Vial 4 | 200 µl, 10 µM | Reverse Primer for Vλ fragment amplification |
| Vial 5 | 200 µl, 10 µM | Forward and reverse primer mix for Ck fragment amplification |
| Vial 6 | 200 µl, 10 µM | Forward and reverse primer mix for Cλ fragment amplification |
| Vial 7 | 200 µl, 10 µM | Forward and reverse primer mix for CH1 fragment amplification |
| Vial 8 | 200 µl, 10 µM | Forward and reverse primer mix for Vk-Ck fragment amplification |
| Vial 9 | 200 µl, 10 µM | Forward and reverse primer mix for Vλ-Cλ fragment amplification |
| Vial 10 | 200 µl, 10 µM | Forward and reverse primer mix for VH-CH1 fragment amplification |
| Vial 11 | 200 µl, 10 µM | Forward and reverse primer mix for Fab fragment amplification |
| pAPD-h/k-Fab and pAPD-h/λ-Fab cloning vector for phage display human Fab library construction | ||
| Vial 12 | 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-h/k-Fab vector or pAPD-h/λ-Fab vector for phage display library construction.
- Cloning Fab candidate into pAPD-h/k-Fab vector or pAPD-h/λ-Fab vector for antibody fragment expression.
