APAF provides services for ‘Functional Proteomics’, the purification of protein bioactives and their assessment for biological activity as indicated by cellular activation or modulation. These capabilities will be underpinned by the purchase and implementation of state-of-the-art protein purification equipment and methodologies, and the development of cell-based high throughput screening assays of biological activity. The cell-based assays will utilize measurement of cellular signal transduction pathways as the index of receptor activation, inhibition or modulation. Development of these assays is at an advanced stage at TGR BioSciences, and it is anticipated that a limited service utilizing these technologies will be available by the end of the calendar year or early 2003. The assays utilize changes in protein states (such as enzymes) as the readout of cellular activation status. The number and further implementation of these assays will be expanded in the coming calendar year.
Development of the protein purification technologies is awaiting delivery of the HPLC equipment. A Gilson HPLC and associated liquid handler have been ordered, and delivery will take place in the next month. This will provide for the separation of proteins by multiple possible resins, and the collection of proteins into 96 well plates. These protein fractions will then be used to activate cultured cells for determination of biological activities.
Advanced liquid handling robots have been purchased from Qiagen to automate the subsampling of protein fractions, the transfer of such fractions to cultured cell lines, and the set-up of the assays of biological activity as determined by the changes in cellular proteins in the cell lysates. Delivery of these robots will occur by the end of November, allowing full implementation by the end of the calendar year.
The Service to be offered, therefore, will be to take a complex mixture of proteins and fractionate the proteins into a more purified form that will be assayed for cell-stimulating/modulating activity, as required by the researcher or organization. Active fractions can then be re-purified further by HPLC and again active fractions monitored through the cell-based assays. This process will be iterated until the sample is pure enough to gain protein sequence data, at which point the sample will be sent to APAF Macquarie Node for such determination. Proteins identified as candidate molecules for cell activation will be further assessed by a variety of techniques, if required, including affinity isolation and cloning.
For more information contact our Adelaide node, located at TGR BioSciences.