Cell-free protein array: Difference between revisions

Revision as of 04:36, 24 May 2022

Revision as of 23:57, 26 July 2022

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The runaway success of [[DNA microarray]]s has generated much enthusiasm for protein microarrays. However, protein microarrays have not quite taken off as expected, even with the necessary tools and know-how from DNA microarrays being in place and ready for adaptation. One major reason is that protein microarrays are much more laborious and technically challenging to construct than DNA microarrays.

The traditional methods of producing protein arrays require the separate ''[[in vivo]]'' expression of hundreds or thousands of proteins, followed by separate purification and immobilization of the proteins on a solid surface. Cell-free protein array technology attempts to simplify protein microarray construction by bypassing the need to express the proteins in [[bacteria]] cells and the subsequent need to purify them. It takes advantage of available [[cell-free protein synthesis]] technology which has demonstrated that protein synthesis can occur without an intact cell as long as cell extracts containing the DNA template, [[transcription (genetics)|transcription]] and [[translation (biology)|translation]] raw materials and machinery are provided.<ref>Katzen, F., G. Chang, et al. (2005). "The past, present and future of cell-free protein synthesis." Trends Biotechnol 23(3): 150–6.</ref> Common sources of cell extracts used in cell-free protein array technology include [[wheat germ]], ''[[Escherichia coli]]'', and rabbit [[reticulocyte]]. Cell extracts from other sources such as [[hyperthermophile]]s, [[hybridoma]]s, [[Xenopus]] [[oocyte]]s, insect, mammalian and human cells have also been used.<ref name="O. Stoevesandt, 2008">He, M., O. Stoevesandt, et al. (2008). "In situ synthesis of protein arrays." Curr Opin Biotechnol 19(1): 4–9.</ref>

The target proteins are synthesized ''[[in situ]]'' on the protein microarray, directly from the DNA template, thus skipping many of the steps in traditional protein microarray production and their accompanying technical limitations. More importantly, the expression of the proteins can be done in parallel, meaning all the proteins can be expressed together in a single reaction. This ability to multiplex protein expression is a major time-saver in the production process.