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Ph.D.-C7C Phage Display Peptide Library Kit v2

Catalog # Concentration Size List Price Quantity Your Price
E8212S 1 set $1,339.00
$1,205.10
Catalog # Size List Price Your Price
E8212S 1 set $1,339.00
$1,205.10
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

The Ph.D.-C7C™ Phage Display Peptide Library Kit v2 contains the Ph.D.-C7C Phage Display Peptide Library, a DYKDDDDK Mouse monoclonal antibody and Protein G Magnetic Beads for a panning control experiment, and enough -96gIII sequencing primer for >50 sequencing reactions. The Ph.D.-C7C Phage Display Peptide Library is based on a combinatorial library of random disulfide looped peptides fused to the N-terminus of a minor coat protein (pIII) of M13 phage. The library consists of ~109 electroporated (i.e., unique) sequences.

  • Ready to use complex phage library (~109 clones)
  • Structurally constrained peptide library
  • Panning against DYKDDDDK Mouse monoclonal antibody yields more reliable epitope results than former target protein (streptavidin)
  • Does not require helper phage for amplification
  • Inherent link between phenotype and genotype allows screening of billions of clones in a single microtiter well or Eppendorf tube
Phage display describes a selection technique in which a library of peptide or protein variants is expressed on the outside of a phage virion, while the genetic material encoding each variant resides on the inside (1-3). This creates a physical linkage between each variant protein sequence and the DNA encoding it, which allows rapid partitioning based on binding affinity to a given target molecule (antibodies, enzymes, cell-surface receptors, etc.) by an in vitro selection process called panning (4). In its simplest form, panning is carried out by incubating a library of phage-displayed peptides on a plate (or bead) coated with the target, washing away the unbound phage, and eluting the specifically bound phage (Figure 1). The eluted phage are then amplified and taken through additional binding/amplification cycles to enrich the pool in favor of binding sequences. After 3–4 rounds, individual clones are characterized by DNA sequencing and binding assays.

The Ph.D.-C7C Phage Display Peptide Library Kit v2 is based on a combinatorial library of random disulfide looped peptides fused to the N-terminus of a minor coat protein (pIII) of M13 phage (5-9). The library is displayed in the form AC-X7-C-GGG, which directly precedes the wild-type pIII sequence. Under neutral conditions disulfide loop formation is favored. Interest in disulfide looped libraries is based on the chance that imposing structural constraints on an unbound ligand may result in a less unfavorable binding entropy upon target binding relative to an unconstrained library (10 -13).  The library consists of ~109 electroporated sequences amplified once to yield approximately 100 copies of each sequence in 10 μl of the supplied phage. 

Displayed Peptide Form: AC-X7-C-GGG-native M13 pIII

Figure 1: Panning with a pentavalent peptide library displayed on pIII.



 
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Ph.D.™-C7C Phage Display Peptide Library Kit v2 E8212-1 -20 1 x 1 set
  Protein G Magnetic Beads S1430V 4 1 x 1 set
Features
  • Control panning experiment does not require blocking step and eliminates spurious plastic binders
  • Control panning experiment with Protein G Magnetic Beads reduces tedium of plate/well washing steps
  • Structurally constrained library may be useful for targets whose native ligands display a surface loop 
  • Ready to use complex phage library matches are theoretically possible for seven randomized positions (207 = 1.28 x 109); inherent link between phenotype and genotype allows screening of billions of clones in a single microtiter well or microfuge tube
  • Does not require helper phage for amplification
  • Discover peptide binders in less than one week with straight-forward molecular biology and sterile culture techniques
  • Supplied -96 gIII Sequencing Primer (500 pmol) allows for >50 sequencing reactions
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

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Notes
  • Recommended long term storage (>30 days) for E. coli K12 ER2738 is -80°C.
  • Protein G Magnetic Beads should be stored at 4°C to prevent damage from freezing. 
  • The original kit (NEB #E8120) and the new version 2 (v2) kit (NEB #E8212) both contain the same Ph.D.-C7C Phage Display Peptide Library component (NEB #E8121).
  • M13 are compatible with most buffer conditions; however, elution buffers may be limited by E. coli culturing conditions needed for amplifications. Amplifications for enriching phage pools may be eliminated if using next generation sequencing strategy. Scan current literature for protocols.
  • Incorporation of a reducing agent in the buffers (e.g. 5 mM DTT), will convert the Ph.D.-C7C Phage Display Peptide Library to a linear library. This feature can also be used to design controls for post-panning binding studies.
  • Supplied -96 gIII Sequencing Primer (500 pmol, 5´-CCCTCATAGTTAGCGTAACG-3 ´) allows for >50 sequencing reactions.
References
  • Sidhu, S.S. (2000). Curr. Opin. Biotechnol.. 11, 610-616.
  • O’Neil, K.T. et al. (1992). Proteins. 14, 509-515.
  • McLafferty, M.A. et al. (1993). Gene. 128, 26-36.
  • Deyle, K. et al . (2017). Acc. Chem. Res. . 50, 1866-1874.
  • Chen, Y. et al. (2006). Nat. Biotech.. 24,
  • Rodi, D.J. and Makowski, L. (1999). Curr. Opin. Biotechnol.. 10, 87-93.
  • Wilson, D.R. and Finlay, B.B. (1998). Can. J. Microbiol.. 44, 313-229.
  • Parmley, S.F. and Smith, G.P. (1988). Gene. 73, 305-318.
  • Scott, J.K. and Smith, G.P. (1990). Science. 249, 386-390.
  • Whaley, S.R. et al. (2000). Nature. 405, 665-668.
  • Noren, K.A. and Noren, C.J. (2001). Methods. 23, 169-178.
  • Rozinov, M.N. and Nolan, G.P (1998). Chem. Biol.. 5, 509-515.
  • Rodi, D.J. et al. (1999). J. Mol. Biol.. 285, 197-203.
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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