Back to Search
Start Over
A phage display vector optimized for the generation of human antibody combinatorial libraries and the molecular cloning of monoclonal antibody fragments
- Source :
- Europe PubMed Central, Scopus-Elsevier
-
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 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.
- Subjects :
- Male
DNA, Complementary
Viral Core Proteins
Genetic Vectors
RNA-Binding Proteins
Middle Aged
Nucleocapsid Proteins
Protein Sorting Signals
Recombinant Proteins
Cell Line
Phagemid vector, Combinatorial antibody library, Phage display, Human monoclonal antibody fragments
Immunoglobulin Fab Fragments
Influenza A Virus, H1N1 Subtype
Lac Operon
Peptide Library
Escherichia coli
Animals
Humans
Cloning, Molecular
Promoter Regions, Genetic
Subjects
Details
- Database :
- OpenAIRE
- Journal :
- Europe PubMed Central, Scopus-Elsevier
- Accession number :
- edsair.pmid.dedup....5c99c49daa3e9af95357479ddd1d6b15