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Title HIGH-THROUGHPUT APTAMER DISCOVERY AND APTAMER INTEGRATION INTO MICROFLUIDIC DEVICES FOR RARE CELL ANALYSIS
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University/Publisher Cornell University
Abstract Precision medicine is the idea where diagnostics and therapeutics are catered to each individual patient to provide personalized care that is optimally effective. For this to be achieved, technologies must exist that extensively examine samples and provide a highly detailed diagnosis for each patient, and processes must exist that can produce personalized drugs that specifically target the patient’s illness. Aptamers are short single stranded nucleic acids that bind to their targets with high affinity and specificity. Aptamers could make a substantial impact toward the goal of precision medicine. However, one of the main challenges preventing aptamers from reaching their potential is the efficient discovery of new high-affinity aptamers. Currently, aptamer selections are very time consuming and expensive, and often do not result in the discovery of a high-quality aptamer. The ability to reliably select aptamers with high affinity and specificity is paramount to the widespread use of aptamers. Consequently, there is great interest in improving selection technology to obtain high-quality aptamers much more rapidly. Toward this effort, we have developed a Microplate-based Enrichment Device Used for the Selection of Aptamers (MEDUSA) that uses affinity microcolumn chromatography. Its versatile 96-well microplate-based design allows this device to be compatible with downstream plate-based processing in aptamer selections, and it lends itself to automation using existing microplate-based liquid-handling systems. MEDUSA is also reconfigurable and is able to operate in serial and/or parallel mode with up to 96 microcolumns. We have demonstrated its use in high-throughput aptamer selections, characterization and optimization of the aptamer selection process, and characterization of previously selected aptamers. More specifically, MEDUSA was used to perform 96 simultaneous tests that determined the optimal target loading on resin to maximize aptamer enrichment for three target proteins, GFP, HSF, and NELF-E. These tests also verified the specificity of aptamers to these three proteins, as well as the non-specific binding of two suspected background binding aptamers. MEDUSA was also used to performed novel RNA aptamer selections to 19 different targets simultaneously. For these selections, a new, more efficient selection strategy was tested that greatly reduced the selection time and reagent consumption. Through the use of MEDUSA, aptamer selections can be optimized and performed in a high-throughput manner, and the success rate of novel aptamer discovery can be drastically improved. In addition to the improvement of novel aptamer discovery, developing valuable applications that use aptamers is of equal importance. An area of study in which aptamers could be of great benefit is cancer. Cancer cells are extremely diverse and contain genetic mutations that allow them to escape the regulatory processes necessary for the healthy function of tissues and organs. Moreover, there are numerous mechanisms for malignancy each with…
Subjects/Keywords Biomedical engineering
Contributors Sondermann, Holger (committeeMember); Kirby, Brian (committeeMember)
Language en
Country of Publication us
Record ID handle:1813/47718
Repository cornell
Date Retrieved
Date Indexed 2019-12-30
Issued Date 2017-01-30 00:00:00

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