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Biopanning is an affinity selection technique which selects for
peptide Peptides (, ) are short chains of amino acids linked by peptide bonds. Long chains of amino acids are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. A ...
s that bind to a given
target Target may refer to: Physical items * Shooting target, used in marksmanship training and various shooting sports ** Bullseye (target), the goal one for which one aims in many of these sports ** Aiming point, in field artillery, fi ...
. All peptide sequences obtained from biopanning using combinatorial peptide libraries have been stored in a special freely available database name
BDB
This technique is often used for the selection of
antibodies An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein used by the immune system to identify and neutralize foreign objects such as pathogenic bacteria and viruses. The antibody recognizes a unique molecule of the ...
too. Biopanning involves 4 major steps for peptide selection. The first step is to have
phage display Phage display is a laboratory technique for the study of protein–protein, protein–peptide, and protein– DNA interactions that uses bacteriophages (viruses that infect bacteria) to connect proteins with the genetic information that encodes ...
libraries prepared. This involves inserting foreign desired
gene In biology, the word gene (from , ; "...Wilhelm Johannsen coined the word gene to describe the Mendelian units of heredity..." meaning ''generation'' or ''birth'' or ''gender'') can have several different meanings. The Mendelian gene is a ba ...
segments into a region of the
bacteriophage A bacteriophage (), also known informally as a ''phage'' (), is a duplodnaviria virus that infects and replicates within bacteria and archaea. The term was derived from "bacteria" and the Greek φαγεῖν ('), meaning "to devour". Bacteri ...
genome, so that the peptide product will be displayed on the surface of the bacteriophage virion. The most often used are genes pIII or pVIII of bacteriophage M13.Smith GP, and Scott JK. ''Libraries of peptides and proteins displayed on filamentous phage''.
Methods in Enzymology ''Methods in Enzymology'' is a book-series of scientific publications focused primarily on research methods in biochemistry by Academic Press, created by Sidney P. Colowick and Nathan O. Kaplan. Content Historically, each volume has centered o ...
. 1993. 217:228-257
The next step is the capturing step. It involves conjugating the phage library to the desired target. This procedure is termed panning. It utilizes the binding interactions so that only specific peptides presented by bacteriophage are bound to the target. For example, selecting antibody presented by bacteriophage with coated
antigen In immunology, an antigen (Ag) is a molecule or molecular structure or any foreign particulate matter or a pollen grain that can bind to a specific antibody or T-cell receptor. The presence of antigens in the body may trigger an immune response. ...
in microtiter plates. The washing step comes after the capturing step to wash away the unbound phages from solid surface. Only the bound phages with strong affinity are kept. The final step involves the elution step where the bound phages are eluted through changing of pH or other environment conditions. The end result is the peptides produced by bacteriophage are specific. The resulting filamentous phages can infect
gram-negative Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the Gram staining method of bacterial differentiation. They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wall ...
bacteria once again to produce phage libraries. The cycle can occur many times resulting with strong affinity binding peptides to the target.


References

{{reflist, 1 Biochemistry methods