Research and development of protein arrays and their applications in proteomics
Before setting up Cambridge Protein Arrays, we have developed the "DNA array to protein array" (DAPA) technology, which creates protein arrays by direct 'printing' from DNA arrays using cell-free protein expression systems.
The template DNA array on one glass slide is sandwiched with a second, protein-capture slide, and cell-free protein synthesis is performed in a narrow gap between the two surfaces. Individual proteins synthesised in parallel from the arrayed DNA become immobilised through interaction with the protein-capturing reagent on the opposite surface. The result is a protein array replica of the DNA array, with good reproducibility and spot quality, as shown:
The same DNA template can be reused several times to generate replicate protein arrays. DAPA makes possible the generation of protein arrays as and when required, in a single in situ reaction. Given a suitable DNA template array, DAPA allows users to generate complex protein microarrays with minimal equipment in just 2 hours. By linking gene sequence data to protein expression, DAPA is an enabling technology for individualised or personalised proteomics. DAPA is in principle scaleable to sub-proteome arrays of hundreds of different proteins and thereafter to full sets of proteomes. We have also developed a method to improve the yield of proteins for which cell free expression is otherwise very poor, by C-terminal fusion of an immunoglobulin light chain Cκ domain, which we have shown to enhance protein production.
He M, Stoevesandt O, Palmer EA, Khan F, Ericsson O, Taussig MJ. Printing protein arrays from DNA arrays. Nat Methods. 2008 5:175-7.
Stoevesandt O, Vetter M, Kastelic D, Palmer EA, He M, Taussig MJ. Cell free expression put on the spot: advances in repeatable protein arraying from DNA (DAPA). N Biotechnol. 2011 28:282-90.
Palmer, E. Liu H, Khan F, Taussig MJ, He M. Enhanced cell-free protein expression by fusion with immunoglobulin Ckappa domain. Protein Science 2006 15: 2842-6.