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Surfaces, Interfaces, and Applications
Glycan-Imprinted Magnetic Nanoparticles-based SELEX for Efficient Screening of Glycoprotein-Binding Aptamers Yanyan Ma, Xinglin Li, Wei Li, and Zhen Liu ACS Appl. Mater. Interfaces, Just Accepted Manuscript • DOI: 10.1021/acsami.8b14441 • Publication Date (Web): 31 Oct 2018 Downloaded from http://pubs.acs.org on November 1, 2018
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ACS Applied Materials & Interfaces
Glycan-Imprinted Magnetic Nanoparticles-based SELEX for Efficient Screening of Glycoprotein-Binding Aptamers Yanyan Ma, Xinglin Li, Wei Li, and Zhen Liu*
State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China
* Corresponding author:
[email protected] 1
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Abstract: Nucleic acid aptamers, as useful alternatives of antibodies, have found a large range of promising applications such as affinity separation and bioassays. The screening of aptamers is critical for their applications. Aptamers are often screened by an in vitro methodology called SELEX (systematic evolution of ligands by exponential enrichment). Although numerous SELEX methods have been established to facilitate the selection, new efficient selection methods are still much needed. Molecularly imprinted polymers (MIPs), which are antibody alternatives at the material level and competitors of aptamers, have not been used as a platform for aptamer selection yet so far. In this study, a glycan-imprinted magnetic nanoparticles (MNPs)-based SELEX was developed to efficiently screen aptamers against glycoproteins. Glycan-imprinted MNPs were used as an affinity interface to bind target glycoprotein and then the target glycoprotein-bound MNPs were used as an affinity substrate for aptamer selection. The glycan-imprinted MNPs were synthesized by a state-of-the-art imprinting approach called boronate affinity controllable oriented surface imprinting (BACOSI). The glycan-imprinted MNPs exhibited high affinity and specificity, and therefore allowed preferential binding towards target glycoproteins while excluding unwanted species. Two representative glycoproteins, including RNase B and TRF, were employed as target glycoproteins, and aptamers with high affinity and specificity towards the two target glycoproteins were screened out in 3 rounds. This method exhibited some merits, such as high affinity, fast speed, and avoiding negative screening. Therefore, the glycan-imprinted MNPs-based SELEX approach holds great values for the efficient screening of high-performance aptamers.
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ACS Applied Materials & Interfaces
Keywords: Aptamer, glycan, glycoprotein, molecular imprinting, molecularly imprinted polymer
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Aptamers, which are RNA or short single-stranded DNA (ssDNA) molecules, exhibit specific binding towards certain target species such as metal ions, small compounds, proteins, viruses, bacteria and cells.1-3 Compared to antibodies, aptamers possess multiple attractive features such as small size, high batch-to-batch reproducibility, ability to sustain reversible denaturation, easy and controllable modification, and lack of immunogenicity.4 Thus, aptamers have great potential as antibody alternatives to be applied in many promising fields such as affinity separation,5,6 disease diagnosis,7,8 disease theraphy9,10 and food security.11 The selection of aptamers is essential for their applications. Nucleic acid aptamers are usually screened from a synthetic oligonucleotide library by SELEX (systematic evolution of ligands by exponential enrichment).12-14 SELEX is an in vitro procedure consist of three major steps: binding, partitioning, and amplification. Classic SELEX, which mainly relied on nitrocellulose membrane filtration13 and affinity separation,14 are usually laborious and time-consuming, requiring up to 12 rounds of selection or a few weeks to obtain aptamers with satisfied affinity. Thus, it is vital to improve the selection efficiency, which is usually measured by the time required or the number of screening rounds. .15 To this end, many new derivatives of SELEX have been developed. For instance, new techniques such as capillary electrophoresis (CE),16,17 magnetic beads,18,19 microfluidics,20,21 microarrays,22,23 and microscopy24,25 have been incorporated and adapted into the SELEX strategy. However, these methods also suffer from several apparent disadvantages. For example, CE-SELEX only collects limited amounts of aptamers at each cycle because of the minute injection volume, thus it is usually difficult to detect the peaks for the aptamer candidate-target complexes.26 And microarray-based SELEX is usually limited by its low capacity (
