Tailoring of Antibody Arrays - Technical Advances Towards Global Proteome Analysis

Abstract: Abstract The number of cancer and autoimmune disease diagnoses increases in our society, which place high demands on the health care system. Being able to tailor treatments for each individual patient is called "Personalized medicine", and could revolutionize the care in the future. This type of medicine requires biomarkers that could be used for diagnosis, prognosis and choice of therapy. In our group, we have developed an antibody microarray discovery tool that can analyze thousands of proteins in clinical samples in one single experiment. By identifying differences in protein expression profiles, biomarkers for various diseases can hopefully be found. To date, antibody microarrays with an overall foot print of < 1 cm2, based on 18x103 µm2 (⌀ ~150 μm) sized spots at a density of ≤ 2,000 spots/cm2, have mainly been produced. Considering the size and complexity of the human proteome, this microarray design will not be able to harbor the number of antibodies required to perform global proteomics, demonstrating the need for novel, miniaturized high-density array designs. Increasing the number of antibodies on the arrays will generate a logistical problem when it comes to purification and immobilization of antibodies, which requires new technical solutions. The aim of this thesis was to develop new methods to produce the next generation of antibody arrays towards global proteome analysis. The first part of this thesis has involved design of miniaturized antibody arrays. The second part has been focused on developing antibodies with a photoreactive property, with the long-term goal to facilitate the logistics when it comes to purification and immobilization of antibodies. In conclusion, new methods to fabricate miniaturized antibody arrays, focusing on both miniaturization and antibody immobilization, for large-scale protein analysis of clinical samples have been designed. In the future, this could lead to better discovery tools for identifying biomarkers for improved diagnosis, prognosis and choice of treatments for various diseases.