Hyperphage M13 KO7ΔpIII (5x 2 ml)

PROGEN Biotechnik GmbH

Functional principle of the Hyperphage System.

1x2 ml (lyoph.)

5x2 ml (lyoph.)

Cat. No. PRHYPE

Prices excl. VAT

Available, delivery time: 1-7 days

€527.00*

Datasheet

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.

References/Publications (37)

Publication

Species

Application

Publication Turrini, E. et al. Molecular engineering of a spheroid-penetrating phage nanovector for photodynamic treatment of colon cancer cells. Cell. Mol. Life Sci. 81, (2024).

Publication Zhu, C. et al. Epitope-Directed Antibody Elicitation by Genetically Encoded Chemical Cross-Linking Reactivity in the Antigen. (2023) doi:10.1021/acscentsci.3c00265.

Publication Costantini, E., Calvaresi, M. & Danielli, A. iScience A modular phage vector platform for targeted photodynamic therapy of Gram-negative bacterial pathogens. ISCIENCE 26, 108032 (2023).

Publication Díaz-Perlas, C. et al. High-affinity peptides developed against calprotectin and their application as synthetic ligands in diagnostic assays. Nat. Commun. 14, 2774 (2023).

Publication Huang, P. L. et al. A bispecific antibody AP203 targeting PD-L1 and CD137 exerts potent antitumor activity without toxicity. J. Transl. Med. 21, 346 (2023).. Commun. 14, 2774 (2023).

Publication Martinez, B. I. et al. Uncovering temporospatial sensitive TBI targeting strategies via in vivo phage display. 5047, (2022).

Publication Bortot, B. et al. Advanced photodynamic therapy with an engineered M13 phage targeting EGFR: Mitochondrial localization and autophagy induction in ovarian cancer cell lines. Free Radic. Biol. Med. 179, 242–251 (2022).

Publication Ballmann, R. et al. ORFeome Phage Display Reveals a Major Immunogenic Epitope on the S2 Subdomain of SARS-CoV-2 Spike Protein. Viruses 14, 1326 (2022).

Publication Dolgikh, V. V. et al. Construction of scFv Antibodies against the Outer Loops of the Microsporidium Nosema bombycis ATP/ADP-Transporters and Selection of the Fragment Efficiently Inhibiting Parasite Growth. Int. J. Mol. Sci. 23, (2022).

Publication Gonschorek, P. et al. Phage Display Selected Cyclic Peptide Inhibitors of Kallikrein-Related Peptidases 5 and 7 and Their in Vivo Delivery to the Skin. J. Med. Chem. (2022).

Publication Zhang, Y. et al. Preparation of a Single-Chain Antibody against Nucleocapsid Protein of Porcine Deltacoronavirus by Phage Display Technology. Viruses 14, (2022).

Publication Seidel-Greven, M. et al. Isolation and light chain shuffling of a Plasmodium falciparum AMA1-specific human monoclonal antibody with growth inhibitory activity. Malar. J. 20, 1–20 (2021).

Publication Carle, V. et al. Generation of a 100-billion cyclic peptide phage display library having a high skeletal diversity. Protein Eng. Des. Sel. 34, 1–9 (2021).

Publication Lawrie, J., Waldrop, S., Morozov, A., Niu, W. & Guo, J. Engineering of a Small Protein Scaffold to Recognize Sulfotyrosine with High Specificity. ACS Chem. Biol. 16, 1508–1517 (2021).

Publication Solemani Zadeh, A., Grässer, A., Dinter, H., Hermes, M. & Schindowski, K. Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries. Methods Protoc. 2, 17 (2019).

Publication Zantow, J. et al. Mining gut microbiome oligopeptides by functional metaproteome display. Sci. Rep. 6, 34337 (2016).

Publication Connor, D. O., Zantow, J., Hust, M., Bier, F. F. & von Nickisch-Rosenegk, M. Identification of Novel Immunogenic Proteins of Neisseria gonorrhoeae by Phage Display. PLoS One 11, e0148986 (2016).

Publication Lai, Y. Da et al. Generation of potent anti-vascular endothelial growth factor neutralizing antibodies from mouse phage display library for cancer therapy. Int. J. Mol. Sci. 17, 1–17 (2016).

Publication Kügler, J. et al. Generation and analysis of the improved human HAL9/10 antibody phage display libraries. BMC Biotechnol. 15, 10 (2015).

Publication Becker, M. et al. Application of M13 phage display for identifying immunogenic proteins from tick (Ixodes scapularis) saliva. BMC Biotechnol. 15, 43 (2015).

Publication Hülseweh, B. et al. Human-like antibodies neutralizing Western equine encephalitis virus. MAbs 6, 718–27 (2014).

Publication Steinwand, M. et al. The influence of antibody fragment format on phage display based affinity maturation of IgG. MAbs 6, 204–18 (2014).

Publication Miethe, S. et al. Development of neutralizing scFv-Fc against botulinum neurotoxin A light chain from a macaque immune library. MAbs 6, 446–59 (2014).

Publication Trott, M. et al. Functional Characterization of Two scFv-Fc Antibodies from an HIV Controller Selected on Soluble HIV-1 Env Complexes: A Neutralizing V3- and a Trimer-Specific gp41 Antibody. PLoS One 9, e97478 (2014).

Publication Hust, M., Frenzel, A., Schirrmann, T. & Dübel, S. Selection of recombinant antibodies from antibody gene libraries. Methods Mol. Biol. 1101, 305–20 (2014).

Publication Zhou, M. et al. Identification of a new epitope for HIV-neutralizing antibodies in the gp41 membrane proximal external region by an Env-tailored phage display library. Eur. J. Immunol. 43, 499–509 (2013).

Publication Rülker, T. et al. Isolation and characterisation of a human-like antibody fragment (scFv) that inactivates VEEV in vitro and in vivo. PLoS One 7, e37242 (2012).

Publication Hust, M., Frenzel, A., Meyer, T., Schirrmann, T. & Dübel, S. Construction of human naive antibody gene libraries. Methods Mol. Biol. 907, 85–107 (2012).

Publication Meyer, T. et al. Identification of immunogenic proteins and generation of antibodies against Salmonella Typhimurium using phage display. BMC Biotechnol. 12, 29 (2012).

Publication Breitling, F. Improving Phage Display Throughput by Using Hyperphage , Miniaturized Titration and pVIII (g8p) ELISA. 7, 197–206 (2010).

Publication Kügler, J. et al. Identification of immunogenic polypeptides from a Mycoplasma hyopneumoniae genome library by phage display. Appl. Microbiol. Biotechnol. 80, 447–458 (2008).

Publication Kirsch, M. I. et al. Development of human antibody fragments using antibody phage display for the detection and diagnosis of Venezuelan equine encephalitis virus (VEEV). BMC Biotechnol. 8, 66 (2008).

Publication Soltes, G. et al. On the influence of vector design on antibody phage display. J. Biotechnol. 127, 626–37 (2007).

Publication Pelat, T. et al. High-affinity, human antibody-like antibody fragment (single-chain variable fragment) neutralizing the lethal factor (LF) of Bacillus anthracis by ... Antimicrob. Agents Chemother. 51, 2758–64 (2007).

Publication Hust, M. et al. Enrichment of open reading frames presented on bacteriophage M13 using Hyperphage. Biotechniques 41, 335–342 (2006).

Publication Broders, O., Breitling, F. & Dübel, S. Hyperphage. Improving antibody presentation in phage display. Methods Mol. Biol. 205, 295–302 (2003).

Downloads

File

Category

Size

Filetype

File PRHYPE packaging protocol

Category Protocol

Size 233.28 KB

Filetype pdf

Datasheet

MSDS

Customer Reviews

0 of 0 reviews

Leave a review!

Share your experiences with other customers.

Login

Your email address

Your password

No reviews found. Share your insights with others.

FAQs

Only vectors coding for a fully functional PIII protein are suitable for the hyperphage system, for example pSEX81.
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.

Contact

Need more product information?