A phage display vector optimized for the generation of human antibody combinatorial libraries and the molecular cloning of monoclonal antibody fragments

作者：Solforosi Laura; Mancini Nicasio; Canducci Filippo; Clementi Nicola; Sautto Giuseppe Andrea; Diotti Roberta Antonia; Clementi Massimo; Burioni Roberto^*

来源：New Microbiologica, 2012, 35(3): 289-294.

• Phagemid vector
• Combinatorial antibody library
• Phage display
• Human monoclonal antibody fragments (mFabs)

A novel phagemid vector, named pCM, was optimized for the cloning and display of antibody fragment (Fab) libraries on the surface of filamentous phage. This vector contains two long DNA %26quot;stuffer%26quot; fragments for easier differentiation of the correctly cut forms of the vector. Moreover, in pCM the fragment at the heavy-chain cloning site contains an acid phosphatase-encoding gene allowing an easy distinction of the Escherichia coli cells containing the unmodified form of the phagemid versus the heavy-chain fragment coding cDNA. In pCM transcription of heavy-chain Fd/gene III and light chain is driven by a single lacZ promoter. The light chain is directed to the periplasm by the ompA signal peptide, whereas the heavy-chain Fd/coat protein III is trafficked by the pelB signal peptide. The phagemid pCM was used to generate a human combinatorial phage display antibody library that allowed the selection of a monoclonal Fab fragment antibody directed against the nucleoprotein (NP) of Influenza A virus.

• 出版日期2012-7