Multifunctional Screening Platform for the Highly Efficient Discovery of Aptamers with High Affinity and Specificity

作者：Hong, Shao-Li; Wan, Ya-Tao; Tang, Man; Pang, Dai-Wen; Zhang, Zhi-Ling^*

来源：Analytical Chemistry, 2017, 89(12): 6535-6542.

DOI：10.1021/acs.analchem.7b00684

• DNA APTAMERS
• MICROFLUIDIC CHANNELS
• NEXT-GENERATION
• SSDNA APTAMER
• CANCER-CELLS
• SELECTION
• SELEX
• PROTEIN
• CHIP
• THERAPEUTICS

Aptamers have attracted much attention as the next generation of affinity reagents. Unfortunately, the selection efficiency remains a critical bottleneck for the widespread application of aptamers. Herein, to accelerate aptamers discovery, a multifunctional microfluidic selection platform was developed, on which the selection efficiency was greatly improved and high-affinity and-specificity aptamers were generated within two round selections. The multifunctional screening platform, precisely manipulating magnetic beads on the micrometer scale, improved selection performance based on microfluidic continuous flow and enhanced the selection process control via in situ monitoring and real-time evaluation. This method could suppress similar to 50-fold nonspecific binding nucleic acids compared to the conventional methods, further eliminate weakly bound nucleic acids within 9 min, and simultaneously perform the negative selection and positive selection. And the selection effectiveness was in situ and real-time monitoring. Three aptamers showed high affinity and specificity toward mucin 1 (MUC1) with dissociation constants (K-d) in nanomolar range (from 22 to 65 nM). Furthermore, the selected aptamer was able to specially label cancer cells and efficiently capture exosomes with 64% capture efficiency. It demonstrated that the multifunctional screening platform was an efficient method to generate high-quality aptamers in a rapid and economic manner.

• 出版日期2017-6-20
• 单位武汉大学