Automated Optimization of Aptamer Selection Buffer Conditions

Open Access
Publisher Website
Google Scholar
Abstract
Optimizing the buffer conditions of the selection of nucleic acid binding species (aptamers), increases the likelihood of producing a target aptamer. Aptamers, with high target affinity and specificity, are often compared to antibodies, as aptamers emerge in the industry as diagnostic and therapeutic tools. The increased demand for aptamers encourages high-throughput aptamer generation. The selection buffer conditions may vary as widely as the selection targets, and therefore buffer optimization is helpful if not required for effective aptamer selections. Such optimization work is time consuming and repetitious, which bodes well for high-throughput applications. To accommodate this, an automated buffer testing protocol has been developed to test target-to-unselected RNA pool binding in the presence of 96 different buffer conditions. The dynamic program may vary the monovalent salt(s) identity, monovalent salt(s) concentration, divalent salt(s) identity, divalent salt concentration, buffer identity, buffer concentration, and pH. The optimized buffer conditions likely increase the probability of a successful selection and therefore promote higher ratios of successful aptamer selections against a variety of targets. Preliminary results show trends with the buffer matrix solutions and lysozyme:unselected pool binding. In general, an inverse relationship between lysozyme binding and monovalent salt concentration is observed. (JALA 2004;9:117-22)