Phage Display procedures (Protocol)

In general, there are 5 steps for phage display technology as below:

STEP1: Construct phage display library

Recombinant DNA technology is used to incorporate foreign cDNA into viral DNA. Different sets of genes are inserted into the genomes of multiple phages. Spliced into genes for a coat protein to display the protein on the outside of phage particles. These individual phages will only display one protein, peptide, or antibody.

Collections of these phages can comprise libraries, such as antibody phage library, protein phage library, or random phage library.

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phage display


STEP2: Binding

These libraries are exposed to selected targets, and only some phages will interact with targets. The target is specific ligands planned to be identified, such as immobilized protein, cell surface protein, or vascular endothelium.

STEP3: Washing

Unbound phages can be washed away, and only those which show an affinity for the receptors are left.

STEP4: Elution

Recovery of the target-bound phage by elution.

STEP5: Amplification

Eluted phages showing specificity are used to infect new host cells for amplification or direct bacterial infection and amplification of the recovered phage.

Back to step 1, repeated cycle 2-3 times for stepwise selection of best binding sequence. After that, you can Enrichment and purification the phage repertoire by precipitation methods to increase the phage titer.

Phage display library screening

About the author

Hello there!
I'm Raphael Hans Lwesya, My true passion lies in the world of phage research and science communication. As a diligent phage researcher and an enthusiastic science communicator, I've founded "www.thephage.xyz," a platform dedicated to unraveling the fascinating universe of bacteriophages – viruses that specifically target microbes. My ultimate mission is to bridge the communication gap between the general public and the often intricate world of scientific concepts. I take pride in simplifying complex ideas, breaking them down into easily understandable pieces, and making cutting-edge phage-related research accessible to a wide audience. Thank you for visiting The Phage blog. If you have got any question or suggestion please drop it as a comment or via [email protected]

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