PD020- Phage Vector fADL-1e -Antibody Design

Technical Description

fADL™-1e is a type 3 phage display vector with a cloning site on the N-terminal side of the full-length gene III protein. Secretion in the periplasm of the fusion protein is driven by the PelB leader peptide, in place of the wild-type gene III protein leader sequence. fADL™-1e derives from the phage vector fd-kan, a vector analogous to fd-tet (1,2) where the tetracycline resistance genes derived from Tn10 have been replaced by the kanamycin resistance determinant of Tn903. Display is usually highly multivalent with limited polyphage production, even in the case of large polypeptides such as scFvs. Implications of multivalent display in phage libraries versus phagemid libraries is discussed here (3). This is the recommended format for peptide libraries which will benefit of the overall improved avidity. A discussion on phage versus phagemid antibody libraries can be found here (3).

fADL™-1e differs from fADL™-1 by the presence of an extra SpeI site in the cloning site and a mutation in protein VIII that improves cross-reactivity with M13 antibodies.

Figue 1. Display of HyHEL-10 scFv by fADL-1. Western blot analysis of phage particles prepared with fADL-1 in the presence of a plasmid supplying gene III protein (Lane A). The same blot with phage particles prepared with fADL-1 containing the HyHEL-10 scFv insert shows the scFv fused to almost all gene III proteins (Lane B). Loading is 3 x 10¹⁰ virions per lane; image was generated by ECL using a gene III protein antibody. Implications of multivalent display in phage libraries versus phagemid libraries is discussed here (3).

Applications

Multivalent Phage display at the N-terminal side of gene III protein of the filamentous bacteriophage

Specifications

General Characteristics:

Plasmid Size: 7993
Leader Peptide: PelB
Cloning Sites: double-SfiI, BglI
Fusion Protein: full length gene III protein
Selection: kanamycin
Replication: oriF1 (60 copies/cells)

Physical characteristics:Add New

Concentration: 0.5 µg/ µl.
Product Size: 10 µg.
Buffer: DNA Conservation Buffer (Tris/HCL 5 mM, EDTA 0.1 mM, pH 8.5, sterile).
Storage Temperature: -20°C.