Screening of the specific aptamer of human CD20 extracellular protein expressed in <i>Escherichia coli</i> by systematic evolution of ligands by exponential enrichment.

Fan CHEN; Fan YANG; Lei GAO; Yue HU; Yun XUE; Jing ZHOU; Jianhua KANG; Wei WANG

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Screening of the specific aptamer of human CD20 extracellular protein expressed in Escherichia coli by systematic evolution of ligands by exponential enrichment.

Author: Fan CHEN ¹ ; Fan YANG ¹ ; Lei GAO ² ; Yue HU ¹ ; Yun XUE ² ; Jing ZHOU ³ ; Jianhua KANG ⁴ ; Wei WANG ⁴
Author Information

1. Life Science School of Hubei University, Wuhan 430062, Hubei, China.
2. School of Medical Technology and Engineering, Henan University of Science and Technology, Luoyang 471003, Henan, China.
3. Wuhan Pulmonary Hospital, Wuhan 430030, Hubei, China.
4. Department of Laboratory Medicine, Hospital of Chang'an Dongguan, Dongguan 523843, Guangdong, China.
Publication Type:Journal Article
Keywords: CD20; aptamer; gene optimization; prokaryotic expression; systematic evolution of ligands by exponential enrichment (SELEX)
MeSH: Humans; Escherichia coli/metabolism*; SELEX Aptamer Technique/methods*; Aptamers, Nucleotide/genetics*; Antigens, CD20/metabolism*; Ligands
From: Chinese Journal of Biotechnology 2025;41(4):1467-1477
CountryChina
Language:Chinese
Abstract: CD20 is a surface marker protein of B-cell lymphoma, and its extracellular region is the target of specific antibodies and drugs. To obtain a cheap and easily modified specific preparation targeting CD20, we optimized the gene of CD20 extracellular region according to codon degeneracy to facilitate its expression in Escherichia coli. The optimized gene was cloned into pGEX-4T-1 vector, and the recombinant vector was transformed into E. coli BL21(DE3) for expression. The purified protein was identified by SDS-PAGE and Western blotting. Systematic evolution of ligands by exponential enrichment (SELEX) was employed to screen the ssDNA aptamer that specifically binds to the fusion protein, and the affinity of the aptamer to CD20 was detected by flow cytometry. Then, the cytotoxicity test was carried out to examine the inhibitory effect of the aptamer on B lymphoma cells. In this study, we established the prokaryotic expression method of CD20 and obtained the aptamer specifically binding to the extracellular region of CD20, which laid a foundation for the development of therapeutic drugs targeting CD20.