A modified SELEX approach to identify DNA aptamers with binding specificity to the major histocompatibility complex presenting ovalbumin model antigen.

Autor:	Lin Y; Department of Medical Research, Taipei Veterans General Hospital 201, Section 2, Shi-Pai Road Taipei 112 Taiwan kfhung@vghtpe.gov.tw +886-2-28712121-7382., Chen CY; Institute of Biomedical Informatics, National Yang Ming Chiao Tung University Taipei Taiwan., Ku YC; Institute of Biomedical Informatics, National Yang Ming Chiao Tung University Taipei Taiwan., Wang LC; Institute of Biomedical Informatics, National Yang Ming Chiao Tung University Taipei Taiwan., Hung CC; School of Computer Science, Georgia Institute of Technology Atlanta GA USA., Lin ZQ; Department of Medical Research, Taipei Veterans General Hospital 201, Section 2, Shi-Pai Road Taipei 112 Taiwan kfhung@vghtpe.gov.tw +886-2-28712121-7382., Chen BH; Department of Medical Research, Taipei Veterans General Hospital 201, Section 2, Shi-Pai Road Taipei 112 Taiwan kfhung@vghtpe.gov.tw +886-2-28712121-7382., Hung JT; Kang Chiao International School Taipei Taiwan., Sun YC; School of Medicine, Tzu-Chi University Hualien Taiwan.; Department of Ophthalmology, Taipei Tzu Chi Hospital, The Buddhist Tzu Chi Medical Foundation New Taipei City Taiwan., Hung KF; Department of Medical Research, Taipei Veterans General Hospital 201, Section 2, Shi-Pai Road Taipei 112 Taiwan kfhung@vghtpe.gov.tw +886-2-28712121-7382.; Department of Dentistry, School of Dentistry, National Yang Ming Chiao Tung University Taipei Taiwan.
Jazyk:	angličtina
Zdroj:	RSC advances [RSC Adv] 2023 Nov 06; Vol. 13 (46), pp. 32681-32693. Date of Electronic Publication: 2023 Nov 06 (Print Publication: 2023).
DOI:	10.1039/d3ra04686a
Abstrakt:	Aptamers have sparked significant interest in cell recognition because of their superior binding specificity and biocompatibility. Cell recognition can be mediated by targeting the major histocompatibility complex (MHC) that presents short peptides derived from intracellular antigens. Although numerous antibodies have demonstrated a specific affinity for the peptide-MHC complex, the number of aptamers that exhibit comparable characteristics is limited. Aptamers are usually selected from large libraries via the Systemic Evolution of Ligands by Exponential Enrichment (SELEX), an iterative process of selection and PCR amplification to enrich a pool of aptamers with high affinity. However, the success rate of aptamer identification is low, possibly due to the presence of complementary sequences or sequences rich in guanine and cytosine that are less accessible for primers. Here, we modified SELEX by employing systemic consecutive selections with minimal PCR amplification. We also modified the analysis by selecting aptamers that were identified in multiple selection rounds rather than those that are highly enriched. Using this approach, we were able to identify two aptamers with binding specificity to cells expressing the ovalbumin alloantigen as a proof of concept. These two aptamers were also discovered among the top 150 abundant candidates, despite not being highly enriched, by performing conventional SELEX. Additionally, we found that highly enriched aptamers tend to contain fractions of the primer sequence and have minimal target affinity. Candidate aptamers are easily missed in the conventional SELEX process. Therefore, our modification for SELEX may facilitate the identification of aptamers for more application in diverse biomedical fields. Significance : we modify the conventional method to improve the efficiency in the identification of the aptamer, a single strand of nucleic acid with binding specificity to the target molecule, showing as a proof of concept that this approach is particularly useful to select aptamers that can selectively bind to cells presenting a particular peptide by the major histocompatibility complex (MHC) on the cell surface. Given that cancer cells may express mutant peptide-MHC complexes that are distinct from those expressed by normal cells, this study sheds light on the potential application of aptamers to cancer cell targeting. Competing Interests: The authors declare that they have no conflicts of interest relating to the subject matter or materials discussed in this article. (This journal is © The Royal Society of Chemistry.)
Databáze:	MEDLINE
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