2008
DOI: 10.1038/nmeth.1210
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Next-generation high-density self-assembling functional protein arrays

Abstract: We report a high-density self assembling protein microarray that displays thousands of proteins, produced and captured in situ from immobilized cDNA templates. Over 1500 unique cDNAs were tested with > 90% success with nearly all proteins displaying yields within 2 fold of the mean, minimal sample variation and good day to day reproducibility. The displayed proteins revealed selective protein interactions. This method will enable various experimental approaches to study protein function in high throughput.

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Cited by 291 publications
(339 citation statements)
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References 13 publications
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“…With conventional protein microarrays it is necessary to purify thousands of in vivo expressed proteins and to spot these purified proteins on a solid surface (5-8). In contrast, in situ synthesis protein microarray technologies simplify protein microarray fabrication by circumventing the steps of in vivo protein expression and purification (9,10,14,15). This streamlining facilitates an increase in the number of target genes that can be assayed, allowing for thousands of protein-encoding plasmids to be spotted at lower cost and in less time.…”
Section: Arabidopsis Thalianamentioning
confidence: 99%
“…With conventional protein microarrays it is necessary to purify thousands of in vivo expressed proteins and to spot these purified proteins on a solid surface (5-8). In contrast, in situ synthesis protein microarray technologies simplify protein microarray fabrication by circumventing the steps of in vivo protein expression and purification (9,10,14,15). This streamlining facilitates an increase in the number of target genes that can be assayed, allowing for thousands of protein-encoding plasmids to be spotted at lower cost and in less time.…”
Section: Arabidopsis Thalianamentioning
confidence: 99%
“…Plasmid DNA was prepared and mixed with NAPPA printing buffer prior to printing as previously described (41). These expression vectors allow the in vitro expression of proteins of interest with a terminal HaloTag.…”
Section: Methodsmentioning
confidence: 99%
“…Protein Array-Our contra capture protein array was built on NAPPA platform that enables the display of thousands of full-length human proteins without the need for labor-intensive protein expression and purification (41). Proteins are synthesized from printed plasmids bearing genes encoding proteins of interest using the in vitro protein expression system.…”
Section: Concept Of Contra Capturementioning
confidence: 99%
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“…In 2004 Ramachandran et al developed a method for transforming spotted DNA arrays into protein arrays, termed nucleic acid programmable protein array (NAPPA) [9,10 ]. Expression-ready biotinylated plasmid DNA is co-spotted on a glass substrate together with a biotinylated antibody.…”
Section: In Vitro Protein Synthesismentioning
confidence: 99%