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

Laura Solforosi, Nicasio Mancini, Filippo Canducci, Nicola Clementi, Giuseppe Andrea Sautto, Roberta Antonia Diotti, Massimo Clementi, Roberto Burioni

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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 "stuffer" 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 pep-tide, 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.

Original languageEnglish
Pages (from-to)289-294
Number of pages6
JournalNew Microbiologica
Volume35
Issue number3
Publication statusPublished - 2012

Fingerprint

Immunoglobulin Fragments
Molecular Cloning
Bacteriophages
Organism Cloning
Monoclonal Antibodies
Light
Periplasm
Immunoglobulin Fab Fragments
Antibodies
Capsid Proteins
Protein Sorting Signals
Acid Phosphatase
Genes
Libraries
Complementary DNA
Escherichia coli
DNA
Influenza A virus NP protein

Keywords

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

ASJC Scopus subject areas

  • Microbiology (medical)

Cite this

A phage display vector optimized for the generation of human antibody combinatorial libraries and the molecular cloning of monoclonal antibody fragments. / Solforosi, Laura; Mancini, Nicasio; Canducci, Filippo; Clementi, Nicola; Sautto, Giuseppe Andrea; Diotti, Roberta Antonia; Clementi, Massimo; Burioni, Roberto.

In: New Microbiologica, Vol. 35, No. 3, 2012, p. 289-294.

Research output: Contribution to journalArticle

Solforosi, Laura ; Mancini, Nicasio ; Canducci, Filippo ; Clementi, Nicola ; Sautto, Giuseppe Andrea ; Diotti, Roberta Antonia ; Clementi, Massimo ; Burioni, Roberto. / A phage display vector optimized for the generation of human antibody combinatorial libraries and the molecular cloning of monoclonal antibody fragments. In: New Microbiologica. 2012 ; Vol. 35, No. 3. pp. 289-294.
@article{093e695cbfe04f76bc85d70df7edcc93,
title = "A phage display vector optimized for the generation of human antibody combinatorial libraries and the molecular cloning of monoclonal antibody fragments",
abstract = "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 {"}stuffer{"} 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 pep-tide, 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.",
keywords = "Combinatorial antibody library, Human monoclonal antibody fragments (mFabs), Phage display, Phagemid vector",
author = "Laura Solforosi and Nicasio Mancini and Filippo Canducci and Nicola Clementi and Sautto, {Giuseppe Andrea} and Diotti, {Roberta Antonia} and Massimo Clementi and Roberto Burioni",
year = "2012",
language = "English",
volume = "35",
pages = "289--294",
journal = "New Microbiologica",
issn = "1121-7138",
publisher = "Luigi Ponzio e figlio Editori",
number = "3",

}

TY - JOUR

T1 - A phage display vector optimized for the generation of human antibody combinatorial libraries and the molecular cloning of monoclonal antibody fragments

AU - Solforosi, Laura

AU - Mancini, Nicasio

AU - Canducci, Filippo

AU - Clementi, Nicola

AU - Sautto, Giuseppe Andrea

AU - Diotti, Roberta Antonia

AU - Clementi, Massimo

AU - Burioni, Roberto

PY - 2012

Y1 - 2012

N2 - 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 "stuffer" 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 pep-tide, 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.

AB - 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 "stuffer" 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 pep-tide, 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.

KW - Combinatorial antibody library

KW - Human monoclonal antibody fragments (mFabs)

KW - Phage display

KW - Phagemid vector

UR - http://www.scopus.com/inward/record.url?scp=84864530741&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864530741&partnerID=8YFLogxK

M3 - Article

VL - 35

SP - 289

EP - 294

JO - New Microbiologica

JF - New Microbiologica

SN - 1121-7138

IS - 3

ER -