Applications of biomolecule entrapment for the sensing and screening of small molecules

In the past 15 years, sol-gel based entrapment of biomolecules has emerged as a practical alternative to less generic 2-dimensional surface immobilization methods for the purpose of creating bioanalytical devices. The research in this thesis aims to expand on the uses of sol-gel entrapment and to apply this immobilization strategy to the emerging field of protein microarrays. The focus is two-fold: (i) to extend the realm of entrapped species to DNA aptamers and demonstrate their use in sensing and enzyme activity reporting capacities; and (ii) to couple the numerous benefits that sol-gel immobilization can provide with protein microarray technology. With regard to the first endeavor, we found that sol-gel derived materials could be used to successfully entrap DNA aptamers. In addition to remaining fully active, selective and sensitive to its substrate, the entrapped aptamer was used to report on enzyme activity in layered silica structures. In the second portion of the thesis the use of sol-gel derived materials for the creation of protein microarrays was investigated. Demonstrations of successful antibody arrays, coupled enzyme arrays based on the co-immobilization of proteins and reporter molecules and coimmobilized kinase-substrate arrays were key points indicating the potential of sol-gel based microarray technology. The success of these projects has advanced the viability for sol-gel derived materials for use with novel biotechnologies and demonstrated that such materials can alleviate many of the limitations encountered with previously utilized techniques.