Nucleic Acid Aptamers (NAAs), are single-stranded ribo- and deoxyribo-oligonucleotide molecules, which fold into complex functional three-dimensional structures. These three-dimensional structures have the ability to bind to target molecules with high affinity and specificity. Nucleic Acid Aptamers are selected through an iterative process called Systematic Evolution of Ligands by Exponential enrichment (SELEX). Here, we implemented Ligand-Guided Selection (LIGS), a variant of SELEX, to elute aptamers against multiple biomarkers expressed in B-cells i.e., CD19 and CD20. During LIGS step, specific aptamers against CD19 and CD20 were competitively eluted using two specific monoclonal antibodies against the same target at 25°C. The eluted multiple libraries were, then sequenced using Illumina high-throughput (HT) DNA sequencing platform. The resulting sequences were analyzed using a novel bioinformatics workflow, which was designed using GALAXY, an online bioinformatics server. Two major sequence families consisting of 18 hit sequences against CD19 and 34 hit sequences against CD20 were identified based on defined enrichment values. The potential hit sequences were subsequently synthesized and screened against biomarker positive and negative cell lines to identify aptamers candidates against respective markers. Collectively, this study establishes LIGS as a state-of-the-art screening technology that can be used to generate highly specific aptamers against multiple receptor-proteins in their native state expressed on one cell population.

Multi-target ligand-guided selection (LIGS) to generate aptamers against B-cell biomarkers

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.