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Selection, identification, and application of DNA aptamers against bovine pregnancy-associated glycoproteins 4

文献类型： 外文期刊

作者： Liu, Changbin 1 ; Lu, Chunxia 3 ; Shi, Guoqing 1 ;

作者机构： 1.Shihezi Univ, Coll Anim Sci & Technol, Shihezi 832003, Peoples R China

2.Xinjiang Acad Agr & Reclamat Sci, Inst Anim Husb & Vet Sci, Shihezi 832000, Peoples R China

3.Yangtze Normal Univ, Life Sci & Technol Inst, Chongqing 408100, Peoples R China

关键词： Pregnancy-associated glycoproteins; SELEX; DNA aptamers; Pregnancy diagnosis; Enzyme-linked aptamer assay

期刊名称：ANALYTICAL AND BIOANALYTICAL CHEMISTRY （ 影响因子：4.142； 五年影响因子：3.863 ）

ISSN： 1618-2642

年卷期： 2020 年 412 卷 18 期

页码：

收录情况： SCI

摘要： The bovine pregnancy-associated glycoproteins (bPAGs) have been widely used as robust markers for early diagnosis of pregnancy in the cattle. The current immune recognition methods for detecting bPAGs are limited and, to a certain extent, are associated with high costs and poor stability of the antibody. Aptamers that are more stable and easily synthesized than antibodies might serve as suitable candidates for the development of rapid detection methods. This paper describes selection and characterization of bPAG4 aptamers and theirs applicability to detect bPAG4 in the serum. In this work, the recombinant bovine pregnancy-associated glycoproteins 4 (bPAG4) with a relative molecular mass of about 48 kDa was successfully expressed in human embryonic kidney 293 (HEK 293) cells. Subsequently, the ssDNA aptamers were selected by systematic evolution of ligands by exponential enrichment (SELEX) using magnetic beads (MB) coated with bPAG4 as target. After 9 rounds of selection, three aptamers with high affinity to bPAG4 (K-d = 11.7 similar to 40.2 nM) were identified. The selected aptamers were successfully used in enzyme-linked aptamer assay (ELAA) to detect bPAG4 at a detection limit of 0.09 ng/mL. Meanwhile, it has been successfully applied for the detection of bPAG4 in serum samples. This work demonstrated that the selected aptamers could be used as promising affinity probes in the development of inexpensive, simple, and sensitive analysis methods for detecting bPAGs.

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