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Phage display cycle. 1) fusion proteins for a viral coat protein + the gene to be evolved (typically an antibody fragment) are expressed in bacteriophage. 2) the library of phage are washed over an immobilised target. 3) the remaining high-affinity binders are used to infect bacteria. 4) the genes encoding the high-affinity binders are isolated.
Smith first described the technique in 1985 when he displayed peptides on filamentous phage by fusing the peptide of interest onto gene III of filamentous phage. [8] He was awarded the 2018 Nobel Prize in Chemistry for this work, sharing his prize with Greg Winter and Frances Arnold.
Biopanning is an affinity selection technique which selects for peptides that bind to a given target. [1] All peptide sequences obtained from biopanning using combinatorial peptide libraries have been stored in a special freely available database named BDB.
Phage Display: A Laboratory Manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. OCLC 43903550, ISBN ...
A genomic library is a collection of overlapping DNA fragments that together make up the total genomic DNA of a single organism. The DNA is stored in a population of identical vectors , each containing a different insert of DNA.
John McCafferty is a British scientist, one of the founders of Cambridge Antibody Technology alongside Sir Gregory Winter and David Chiswell. He is well known as one of the inventors of scFv antibody fragment phage display, [1] a technology that revolutionised the monoclonal antibody drug discovery.
Infection begins when the gp9 tailspike of the P22 phage binds to the O-antigen lipopolysaccharide on the surface of Salmonella typhimurium host. [1] The virion's tail fiber protein has endorhamnosidase activity, which cleaves the O-antigen chain. [3] Upon infection, P22 can enter either a lytic or lysogenic growth pathway. [1]
The lytic protein, P5, is contained between the P8 nucleocapsid shell and the viral envelope. The completed phage progeny remain in the cytosol until sufficient levels of the lytic protein P5 degrade the host cell wall. The cytosol then bursts forth, disrupting the outer membrane, releasing the phage. The bacterium is killed by this lysis.