Phage Display
A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".
Deadline for manuscript submissions: closed (29 February 2012) | Viewed by 65665
Special Issue Editors
Special Issue Information
Dear Colleagues,
Antibodies have proven to be one of the most important classes of biological molecules with widespread use as research, diagnostic and therapeutic reagents, due to their unique and high affinity binding to a diverse range of targets. In vitro display technology, such as the display of antibody fragments on the surface of M13, fd and f1 filamentous bacteriophage (phage display), offers a highly powerful method capable of generating antibodies to any desired target with specificity and affinity that is not achievable using established hybridoma approaches [1].
The success or failure of in vitro display is dependent on a number of variables including;
- Size and diversity of immunoglobulin gene library
- Quality and purity of target antigen
- Complexity of target
- Biopanning strategies
Immunoglobulin gene libraries can be composed of natural antibody sequences isolated from either naïve or immunised antibody-producing cells, or can be synthesised to contain random complementarity-determining sequences, allowing isolation of non-naturally occurring binders. Antigens for panning are commonly purified native or recombinant proteins, or whole cells or viruses, but may also be non-protein samples such as lipids or carbohydrates.
The isolation of antibodies against a given target antigen is relatively straight forward, and three to four iterations (rounds) of biopanning generally result in the isolation of one or more antibodies to different epitopes located on the target. However there are more complex scenarios for isolation of antibodies against targets, which require designing specific strategies for biopanning, often incorporating combinations of negative selection steps, to deplete the repertoire of irrelevant antibodies. Such examples include isolation of antibodies against viral antigens that are able to discriminate between closely related subtypes [2], or panning against whole cells where the desired target is only a fraction of the available epitopes present on the cell.
Phage display is ideal for such complex scenarios as experimental conditions can easily be altered such as stringency of selection or negative depletion. Additionally, isolated antibodies can easily be mutagenised for affinity maturation.
Prof. Dr. Stephen Mahler
Dr. Martina Jones
Guest Editors
References
1. Smith, G.P.; Petrenko, V.A. Phage Display. Chem. Rev. 1997, 97, 391–410.
2. Bradbury, A.R.M.; Sidhu, S.; Dubel, S.; McCafferty, J. Beyond natural antibodies: the power of in vitro display technologies. Nat. Biotechnol. 2011, 29, 245–254
Keywords
- antibodies
- biopanning
- phage display
- subtractive selection
- whole cell panning
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