Cell-free protein array: Difference between revisions

Revision as of 01:49, 9 January 2013

Revision as of 16:11, 12 April 2013

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==Methods of synthesis==

=== ''In situ'' methods ===

In the ''in situ'' method, protein synthesis is carried out on a protein array surface that is pre-coated with a protein-capturing reagent or [[antibody]]. Once the newly synthesized proteins are released from the [[ribosome]], the [[protein tag|tag sequence]] that is also synthesized at the [[N-terminus|N-]] or [[C-terminus]] of each nascent protein will be bound by the capture reagent or antibody, thus immobilizing the proteins to form an array. Commonly used tags include [[Polyhistidine-tag|polyhistidine]] (His)6 and [[Glutathione-S-transferase|glutathione s-transferase]] (GST).

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==Applications==

*'''Protein interactions''': To screen for [[protein–protein interactions]]<ref name="Ramachandran, N. 2004"/> and protein interactions with other molecules such as [[metabolite]]s, [[lipid]]s, DNA and small molecules.<ref>He, M. and M. W. Wang (2007). "Arraying proteins by cell-free synthesis." Biomol Eng 24(4): 375–80.</ref>

*'''Enzyme inhibition assay''':<ref name="Angenendt, P. 2004"/>: For high throughput drug candidate screening and to discover novel [[enzyme]]s for use in [[biotechnology]].

*'''Screening antibody specificity'''<ref>He, M. and M. J. Taussig (2003). "DiscernArray technology: a cell-free method for the generation of protein arrays from PCR DNA." J Immunol Methods 274(1–2): 265–70.</ref>