Beschreibung
An Effective Tool for the Isolation of Recombinant Antibodies, Proteins or Peptides from Hyperphage-Packed Libraries.
Advantage:
- increases panning efficiency
- allows panning with reduced amount of panning antigen
- identifies high and low affinity binders
Introduction:
A helper phage technology ("Hyperphage System") was developed by Rondot et al. (Nature Biotechnology 19:75-81, 2001). The Hyperphage System allows to improve antibody presentation in phage display by increasing the number of antibodies displayed per phage particle (up to 5 vs. 0.01) and thereby the system offers great advantage in the fields of functional gene analysis and proteomics. Panning of phages can be performed with small amounts of antigen and higher efficiency. For example, the application of universal libraries for antibody isolation can be improved by employing panning of hyperphage-packed libraries on blots of protein spots after 2-dimensional gel electrophoresis.
More Applications:
Large numbers of open reading frames (ORFs) can be analyzed by panning against synthetic membrane- bound peptide epitopes. In cancer research, the hyperphage-packed library could be a tool to discover new tumour markers by panning against cellular surfaces.
The Hyperphage System:
Hyperphages carry a deletion in the pIII gene. They are generated by an E. coli packaging cell line producing functional pIII which is used to package a phage genome with a pIII deletion. The resulting hyperphages carry functional pIII on their surface but lack the pIII gene in their genome. These hyperphages can then be used to infect bacteria with a phagemid library. Each of the resulting display phages carries several copies of the antibody or peptide on its surface, thus dramatically increasing panning efficiency.
Example with pSEX Phagemid Antibody Gene Library:
Antigen binding was enhanced by more than two orders of magnitude by using hyperphage. Further, since the antibody carrying plasmid (phagemid) encodes a protease cleavage site between pIII and scFv fragment, the hyperphage-packed library can be eluted by protease treatment, allowing to elute the highest affinity binders, plus restoring wild-type infectivity phenotype to optimize the recovery of the antibody gene of interest.
Verwandte Produkte
anti-M13/fd/F1 Filamentous Phages
The antibody binds to pVIII major coat protein of filamentous phages of the fd, F1 and M13 group. Suitable for the identification of recombinant phages in antigen and antibody-phage display systems and phage ELISA.anti-M13/fd/F1 Filamentous Phages
The antibody binds to pVIII major coat protein of filamentous phages of the fd, F1 and M13 group. Suitable for the identification of recombinant phages in antigen and antibody-phage display systems and phage ELISA.pOPE 101 Expression Vector
Cassette vector for the expression of functional recombinant single-chain Fv antibody fragments in Escherichia coli.Cloning of heavy and light chain variable domain genes of Fv antibody fragments selected from hybridomas, single B-lymphocytes or phage display libraries.Hyperphage M13 K07ΔpIII (1x2mL)
Provides helper phage function in packaging a common phage display phagemid. Infection of bacteria via pIII.Phage Titration ELISA
Kit allows fast titration of M13 particles by immunological recognition of protein VIII. Contains standard M13 titrated by electron microscopical method.anti-M13/fd/F1 Filamentous Phages Biotin
The antibody binds to pVIII major coat protein of filamentous phages of the fd, F1 and M13 group. Suitable for the identification of recombinant phages in antigen and antibody-phage systems and phage ELISA.pSEX81 Surface Expression Phagemid Vector
Phagemid cassette vector for the expression of functional recombinant single-chain Fv antibody - pIII fusion proteins on the surface of M13 filamentous phage.Mouse IgG Library Primer Set
PCR oligonucleotide primer sets consisting of 25 primers for amplification and 25 primers for cloning of mouse IgG heavy chain variable domain coding regions for generating universal or antibody-specific scFv phage-display libraries.Human IgG / IgM Library Primer Set
PCR oligonucleotide primer sets consisting of 16 primers for amplification and 16 primers for cloning of human IgG and IgM heavy chain variable domain coding regions for generating universal, patient or antibody-specific scFv phage-display libraries.