Array-Based Discovery of Aptamer Pairs
Abstract
Affinity reagent pairs that recognize distinct epitopes on a target protein can greatly improve the sensitivity and specificity of molecular detection. Importantly, such pairs can be conjugated to generate reagents that achieve two site bidentate target recognition, with affinities greatly exceeding either monovalent component. DNA aptamers are especially well-suited for such constructs, because they can be linked via standard synthesis techniques without requiring chemical conjugation. Unfortunately, aptamer pairs are difficult to generate, primarily because conventional selection methods preferentially yield aptamers that recognize a dominant hot spot epitope. Our array-based discovery platform for multivalent aptamers (AD-MAP) overcomes this problem to achieve efficient discovery of aptamer pairs. We use microfluidic selection and high-throughput sequencing to obtain an enriched pool of aptamer sequences. Next, we synthesize a custom array based on these sequences, and perform parallel affinity measurements to identify the highest-affinity aptamer for the target protein.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 11, 2014
- Accession Number
- AD1027869
Entities
People
- H. T. Soh
- James A. Thomson
- Jeff Nie
- Michael Eisenstein
- Minseon Cho
- Monte J. Radeke
- Peter J. Coffey
- Ron Stewart
- Seung S. Oh
Organizations
- University of California, Santa Barbara