Speaker Abstract RCMM 2017: Dr. Dayangku Haslina Pangiran Razali

Full name: Dr. Dk. Haslina binti Pg. Razali

Designation: Senior Research Scientist

Institution: Sengenics Sdn Bhd

Title:

Development and Application of Full-Length, Correctly Folded Human Protein Microarrays: Drug Selectivity Screening and Diagnostic Biomarker Discovery

Abstract:

Recombinant protein expression is a fundamental technique that underpins much of diagnostics, drug discovery, vaccine development and pure research for elucidating mechanisms of disease development and progression. However, robust, reproducible, high-throughput production of correctly folded and functional full-length human proteins in sufficient scale for downstream functional assays is fraught with difficulties, typically resulting from low-yields of recombinant proteins and individually-optimised purification strategies. We have developed a novel methodology that results in high-throughput expression of correctly folded proteins by using the biotin carboxyl carrier protein (BCCP) as a folding marker and solubility enhancer. The BCCP-protein fusions are capable of being biotinylated either in vivo or in vitro, allowing the use of highly specific biotin-streptavidin interaction for surface capture. We have used this innovative surface technology in the development of a novel, protein microarray platform in which individually-purified BCCP-tagged proteins are immobilised onto purpose-designed surfaces such that they retain folded structure and function and behave in miniaturised, highly multiplexed quantitative assays as if they are in free solution. We have utilised this protein microarray platform in various medical and therapeutic proteomics applications, including autoantibody biomarker profiling, stratification of patients in clinical settings, characterisation of the immunomodulatory properties of antibodies, identification of novel drug targets, as well as predictive toxicology. This presentation will describe how these applications have successfully facilitated the identification, validation and discovery of new biomarkers in cancer, autoimmune and infectious diseases.