New 3D Black Substrate for Protein Microarrays with Improved Dynamic Range
Poster Jan 03, 2006
Markus Hollas, Eric Jallerat, Karl Pflanz, Inka Praulich, Johanna Walter
IntroductionSurge in proteomics research has accelerated the development of new protein arrays as an indispensable tool to study high density protein interaction. The most important factor that determines the performance of protein microarrays is the substrate on which the proteins are immobilised. The microarray substrate, used either in an interaction or capture mode has to provide several key functions. High binding capacity for any proteins without changing their biologically active conformation as well as a low auto fluorescence in order to generate a high signal to noise ratio are essential. This enables the detection of the interaction or capture event within a widedynamic range.
A Microarray slide combining the benefits of increased surface area of 3D cellulose nitrate with the excellent low auto fluorescence of a black substrate has been developed and evaluated. The 3D structure was designed to offer a high binding affinity for proteins and to better stabilize their active conformation even in presence of denaturing temperature conditions. A competitive study analysing the background fluorescence and the detectable concentration range of a chosen antigen is presented. An Affymetrix Spotting roboter 417 arrayer was used for protein dotting. The analysis is based on fluorescence scanning experiments with an Affymetrix 428 array scanner at a gain of 60. Data analysis was made with ImaGene Premium 5.5.3 by Biodiscovery Inc.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE