A comparative study of aptamer isolation by conventional and microfluidic strategies.
A comparative study of aptamer isolation by conventional and microfluidic strategies.
复制标题
DOI：
10.1039/d2an01767a
复制
发表时间：
2023-02-13
期刊：
影响因子：
4.2
通讯作者：
Lin, Qiao
中科院分区：
文献类型：
--
作者：
Meng, Xin;Wen, Kechun;Citartan, Marimuthu;Lin, Qiao
适配体是一种单链寡核苷酸分子，具有高亲和力和特异性，可以与广泛的靶分子结合。指数富集配体的系统进化方法（SELEX）在从随机寡核苷酸库中分离适配体中起着至关重要的作用。迄今为止，SELEX工艺的重大修改和改进已经实现，产生了各种形式的SELEX，从传统的SELEX到基于微流体的全芯片SELEX。虽然全芯片SELEX通常被认为优于传统的SELEX，但尚未对两种方法进行结论性比较。在此，我们对三种SELEX策略进行了比较研究，包括传统的琼脂糖珠分割、微流控亲和选择和完全集成的微流控亲和选择和PCR扩增。使用免疫球蛋白E （IgE）作为模型靶分子，我们比较了这些策略在SELEX过程中每个步骤的时间和成本，包括亲和选择、扩增和寡核苷酸调节。对富集池中的靶标结合寡核苷酸进行测序和比较，以评估SELEX策略的相对有效性。我们表明，微流控策略比传统的SELEX更具时间和成本效益。
Aptamers are single-stranded oligonucleotide molecules that bind with high affinity and specificity to a wide range of target molecules. The method of systematic evolution of ligands by exponential enrichment (SELEX) plays an essential role in the isolation of aptamers from a randomized oligonucleotide library. To date, significant modifications and improvements of the SELEX process have been achieved, engendering various forms of SELEX from conventional SELEX to microfluidics-based full-chip SELEX. While full-chip SELEX is generally considered advantageous over conventional SELEX, there has not yet been a conclusive comparison between the methods. Herein, we present a comparative study of three SELEX strategies for aptamer isolation, including those using conventional agarose bead-based partitioning, microfluidic affinity selection, and fully integrated microfluidic affinity selection and PCR amplification. Using immunoglobulin E (IgE) as a model target molecule, we compare these strategies in terms of the time and cost for each step of the SELEX process including affinity selection, amplification, and oligonucleotide conditioning. Target-binding oligonucleotides in the enriched pools are sequenced and compared to assess the relative efficacy of the SELEX strategies. We show that the microfluidic strategies are more time- and cost-efficient than conventional SELEX.
登录
查看更多内容
影响因子：
17.1
作者：
Qu H;Csordas AT;Wang J;Oh SS;Eisenstein MS;Soh HT
通讯作者：
Soh HT
影响因子：
3.9
作者：
Olsen TR;Tapia-Alveal C;Yang KA;Zhang X;Pereira LJ;Farmakidis N;Pei R;Stojanovic MN;Lin Q
通讯作者：
Lin Q
影响因子：
4
作者：
Macdonald, Joanna;Houghton, Patrick;Shigdar, Sarah
通讯作者：
Shigdar, Sarah
影响因子：
2.9
作者：
Minagawa, Hirotaka;Kataoka, Yuka;Waga, Iwao
通讯作者：
Waga, Iwao
影响因子：
4.6
作者：
Kim, Jinho;Olsen, Timothy R.;Lin, Qiao
通讯作者：
Lin, Qiao