pADL-43

pADL-43

Technical Description

pADL™-43 is a phagemid vector designed for phage display on the C-terminal domain of the protein III of the filamentous bacteriophage M13 or equivalent. This vector contains a PelB leader sequence for expression in the periplasm, a double-SfiI cloning site to introduce scFvs or Fab fragments, a HIS tag for purification, a Myc tag for detection and an amber codon located before the C-terminal domain of the gene III sequence. On non-suppressive bacterial strains, free scFvs or Fab fragments are produced in the periplasm where they can be assayed for binding or purified for further testing. Expression of the fusion is under the control of a lac promoter that has been adjusted at a level comparable to the widely used phagemid pComb3X (1).

Figure 1. Fab display with pADL-43. The binding of a Fab displayed on either pADL-43 or pADL-23c is analyzed by phage ELISA.  Display on the truncated pIII through pADL-43 leads to similar binding to a high affinity epitope (Kappa chain, left panel) and a better binding to a low affinity epitope (a peptide in the paratope, right panel).

Applications

• Phage display at the C-terminal domain of gene III protein of the filamentous bacteriophage.

• Free scFv or Fab expression.

For research use only; not intended for any animal or human therapeutic or diagnostic use.

Specifications

General Characteristics:

Plasmid Size: 3280
Promoter: lac promoter
Leader Peptide: PelB
Cloning Site: double-BglI/SfiI, NotI-SpeI
Purification: HIS tag
Detection: Myc tag
Fusion Protein: C-terminal domain gene III protein and conditional Amber stop codon
Selection: ampicillin
Replication: oriF1, pMB1

Physical Characteristics:

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.

Quality Control & Certification of Analysis

Product Size:

Digestion by EcoRI and NheI generates 2 fragments of 2.5 kb and 0.7 kb.

Product Sequence:

Complete vector sequence is analyzed.

Certification:

Product meets all specifications.

References

1. Rader C. (2023). The pComb3 Phagemid Family of Phage Display Vectors. Cold Spring Harb Protoc. Jul 17. doi: 10.1101/pdb.over107756. Epub ahead of print.