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Hyperphage M13 KO7ΔpIII (5x 2 ml)

Cat. No: PRHYPE

Available, delivery time 1-7 days

$630.00*

Key Features
  • Helper phage for phage display
  • Tool for isolation of recombinant antibodies, proteins and peptides
  • Increased panning efficiency
  • Panning with reduced amount of panning antigen
  • Identification of high and low affinity binders 
Product description
Quantity 5x2 ml (lyoph.)
Purification PEG precipitation
Storage 2-8°C
Intended use Research use only
Reconstitution Reconstitute each vial with 2 ml H2O dist.
Application Provides helper phage function in packaging a common phage display phagemid (for example pSEX, pHen2)
Specificity Infection of bacteria via pIII
Working dilution The concentration of hyperphage particles per ml after reconstitution is indicated on the label. For infection use a MOI of 20
Stability After reconstitution use immediately
Synonym Hyperphage
Background

An Effective Tool for the Isolation of Recombinant Antibodies, Proteins or Peptides from Hyperphage-Packed Libraries.

Advantages

  • 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.

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FAQs
Only vectors coding for a fully functional PIII protein are suitable for the hyperphage system, for example pSEX81, pComb3/pComb3X or similar vectors.
According to the literature pCANTAB vector is compatible with hyperphage, although with lower efficiency as for example pSEX81 (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1866265/).

TG1 is compatible with hyperphage and the classical strain for phage display. It is very suitable for panning (one panning round per day).
For the last panning round XL1-Blue MRF’ may be used, since the plasmid stability, quality and yield is higher with this strain.
Alternatively to TG1 or XL1-Blue MRF’, ER2783 is also suitable for library cloning and packaging.

After reconstitution, the hyperphage can be kept at 2-8°C for several days, however, the infection potential will decrease gradually.
Hyperphage stored at 2-8°C for several weeks (up to 2 months) were still active, however, the pfu was more than 50% decreased.
For long term storage it is possible to add 20% glycerol and store the hyperphage at -80°C. This will also lead to a decrease in activity.

10 mM Tris-HCl (pH 7.5), 20 mM NaCl, 2 mM Na-EDTA
The concentration of our hyperphage is lot dependent. We try to keep the concentration > 1E+12 particles/ml. The specific concentration is indicated on the product label. Please contact us to ask for the concentration of your specific lot or the current lot in stock.
Please be informed that the packaging cell line has been patented (i.e. it is proprietary) and is unfortunately not available publicly.
Therefore it is not possible to amplify hyperphage in your lab.

anti-M13/fd/F1 Filamentous Phages mouse monoclonal, B62-FE2, lyophilized, purified, 100 µg
Cat. No: 61097
Quantity: 100 µg
  • Purified, lyophilized
  • Mouse monoclonal
  • Suitable for ELISA, phage display, IEM and WB
  • Reacts with filamentous phages F1, filamentous phages fd and filamentous phages M13
  • Isotype: IgG2b

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  • Tool for isolation of recombinant antibodies, proteins and peptides
  • Increased panning efficiency
  • Panning with reduced amount of panning antigen
  • Identification of high and low affinity binders 

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