Has anyone else used the M13KE vector for an scFv phage antibody library_

2015-01-13

azim58 - Has anyone else used the M13KE vector for an scFv phage antibody
library?

Yes!
see note at 1415

**Phage** display: Concept, innovations, applications and future

We have screened phage display libraries made in M13KE vector
in which random peptides are fused to the minor coat protein pIII
Note: in this sentence they were just referring to the display of peptides

http://www.nature.com/onc/journal/v29/n32/full/onc2010195a.html

Peptide mimotopes recognized by antibodies cetuximab and matuzumab induce
a functionally equivalent anti-EGFR immune response

The peptides were selected from phage-displayed random peptide libraries
by delayed infectivity panning (Benhar //et al.//, 2000) using
Escherichia coli cells that carry single-chain Fv (scFv) antibody
fragments of cetuximab and matuzumab on their surface.

The Ties that Bind: Peptide Display Technology
(this site has quite a nice overview of different phage technologies and
products)
Immunologists should take note of Amersham Pharmacia's Recombinant Phage
Antibody System (RPAS)--an integrated modular system designed for cloning
recombinant antibody fragments from mice or cell culture and expressing
them in bacteria via phage display. The components are available as
individual modules and support products that provide a systematic
approach for cloning, expressing, detecting, and purifying single-chain
fragment variable (ScFv) antibodies. The core system consists of the
following three modules: Mouse ScFv, Expression, and Detection Modules.

Read more: The Ties That Bind: Peptide Display Technology - The Scientist
- Magazine of the Life Sciences
http://www.the-scientist.com/article/display/18459/#ixzz1i3PvsX8j

1415
Finally! Some direct evidence that someone else has used the M13KE vector
from New England Biolabs with scFv! This is in a patent document.
http://www.freepatentsonline.com/y2008/0206784.html

Construction of Plasmid for Expression of pIII Fused with HEL-Binding
scFv and Confirmation of its Expression

M13KE (manufactured by NEW ENGLAND BIOLABS.) is used for pIII fusion
protein expression. HEL-binding scFv (SEQ ID NO: 97 or 98) is inserted
into the multicloning site of the M13KE. This plasmid is inserted into
Acc65I/EagI according to the technical bulletin of the company. The
resulting plasmid for expression is designated as pM13-HpIII. E. coli is
transformed with pM13-GIII by electroporation according to the technical
bulletin to express a phage displaying pIII fused with HEL-binding scFv.

Inventors of this patent
Shiotsuka, Hidenori (Kawasaki-shi, JP)
Imamura, Takeshi (Chigasaki-shi, JP)

These inventors were also found as authors of this paper
Direct Immobilization of Gold-Binding Antibody Fragments for Immunosensor
Applications
The corresponding author of this paper is
ibii.takahisa@canon.co.jp

Here is my e-mail to Ibii Takahisa

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