{"title":"针对牛妊娠相关糖蛋白的适配体的结构优化、表征和结合机制评估，以及将其应用于利用铁基金属有机框架作为过氧化物酶模拟标记建立比色适配体传感器的研究","authors":"Chunxia Lu, Jiaxiang Qin, Shijia Wu, ZhenLiang Zhang, Zonggui Tang, Changbin Liu","doi":"10.1007/s00604-024-06775-6","DOIUrl":null,"url":null,"abstract":"<div><p> A truncated aptamer (designated A24-3) was identified that specifically binds to bovine pregnancy-associated glycoproteins (bPAG9) with a low dissociation constant (2.04 nM) through two truncation approaches. Circular dichroism spectroscopy indicated that A24-3 formed parallel G-quadruplexes, which was subsequently confirmed using nuclear magnetic resonance (NMR) spectroscopy. Furthermore, a molecular dynamics simulation was employed to investigate the recognition mechanism of A24-3 and bPAG9. Interaction analysis showed that A24-3 folded into a parallel G-quadruplex structure with three G-tetrads, primarily through numerous hydrogen bonds and hydrophobic and π-π interactions. Finally, a novel colorimetric aptasensor was developed for detecting bPAG9 using A24-3 and an Fe-based metal–organic framework as target recognition elements and enzyme mimics, respectively. The method demonstrated a broad detection range from 0.5 to 50 ng/mL, with a low detection limit of 0.03 ng mL<sup>−1</sup>, and exhibited a good recovery (91.0–102%) for bPAG9-spiked serum samples. Additionally, the aptasensor was successfully applied to detecting the pregnancy-specific biomarker bPAGs in serum samples.</p><h3>Graphical Abstract</h3>\n<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>","PeriodicalId":705,"journal":{"name":"Microchimica Acta","volume":"191 11","pages":""},"PeriodicalIF":5.3000,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Structural optimization, characterization, and evaluation of binding mechanism of aptamers against bovine pregnancy-associated glycoproteins and their application in establishment of a colorimetric aptasensor using Fe-based metal-organic framework as peroxidase mimic tags\",\"authors\":\"Chunxia Lu, Jiaxiang Qin, Shijia Wu, ZhenLiang Zhang, Zonggui Tang, Changbin Liu\",\"doi\":\"10.1007/s00604-024-06775-6\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p> A truncated aptamer (designated A24-3) was identified that specifically binds to bovine pregnancy-associated glycoproteins (bPAG9) with a low dissociation constant (2.04 nM) through two truncation approaches. Circular dichroism spectroscopy indicated that A24-3 formed parallel G-quadruplexes, which was subsequently confirmed using nuclear magnetic resonance (NMR) spectroscopy. Furthermore, a molecular dynamics simulation was employed to investigate the recognition mechanism of A24-3 and bPAG9. Interaction analysis showed that A24-3 folded into a parallel G-quadruplex structure with three G-tetrads, primarily through numerous hydrogen bonds and hydrophobic and π-π interactions. Finally, a novel colorimetric aptasensor was developed for detecting bPAG9 using A24-3 and an Fe-based metal–organic framework as target recognition elements and enzyme mimics, respectively. The method demonstrated a broad detection range from 0.5 to 50 ng/mL, with a low detection limit of 0.03 ng mL<sup>−1</sup>, and exhibited a good recovery (91.0–102%) for bPAG9-spiked serum samples. Additionally, the aptasensor was successfully applied to detecting the pregnancy-specific biomarker bPAGs in serum samples.</p><h3>Graphical Abstract</h3>\\n<div><figure><div><div><picture><source><img></source></picture></div></div></figure></div></div>\",\"PeriodicalId\":705,\"journal\":{\"name\":\"Microchimica Acta\",\"volume\":\"191 11\",\"pages\":\"\"},\"PeriodicalIF\":5.3000,\"publicationDate\":\"2024-10-29\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Microchimica Acta\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://link.springer.com/article/10.1007/s00604-024-06775-6\",\"RegionNum\":2,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Microchimica Acta","FirstCategoryId":"92","ListUrlMain":"https://link.springer.com/article/10.1007/s00604-024-06775-6","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
引用次数: 0
摘要
通过两种截短方法鉴定出了一种截短的适配体（命名为 A24-3），它能以较低的解离常数（2.04 nM）与牛妊娠相关糖蛋白（bPAG9）特异性结合。圆二色性光谱显示 A24-3 形成了平行的 G 型四联体，随后核磁共振（NMR）光谱证实了这一点。此外，还利用分子动力学模拟研究了 A24-3 和 bPAG9 的识别机制。相互作用分析表明，A24-3 主要通过大量氢键、疏水作用和 π-π 相互作用折叠成具有三个 G 四叉的平行 G 四叉结构。最后，以 A24-3 和铁基金属有机框架分别作为目标识别元件和酶模拟物，开发了一种新型比色适配传感器，用于检测 bPAG9。该方法的检测范围为 0.5 至 50 ng/mL，检测限低至 0.03 ng mL-1，对添加了 bPAG9 的血清样品具有良好的回收率（91.0-102%）。此外，该传感器还成功地应用于检测血清样本中的妊娠特异性生物标志物bPAGs。
Structural optimization, characterization, and evaluation of binding mechanism of aptamers against bovine pregnancy-associated glycoproteins and their application in establishment of a colorimetric aptasensor using Fe-based metal-organic framework as peroxidase mimic tags
A truncated aptamer (designated A24-3) was identified that specifically binds to bovine pregnancy-associated glycoproteins (bPAG9) with a low dissociation constant (2.04 nM) through two truncation approaches. Circular dichroism spectroscopy indicated that A24-3 formed parallel G-quadruplexes, which was subsequently confirmed using nuclear magnetic resonance (NMR) spectroscopy. Furthermore, a molecular dynamics simulation was employed to investigate the recognition mechanism of A24-3 and bPAG9. Interaction analysis showed that A24-3 folded into a parallel G-quadruplex structure with three G-tetrads, primarily through numerous hydrogen bonds and hydrophobic and π-π interactions. Finally, a novel colorimetric aptasensor was developed for detecting bPAG9 using A24-3 and an Fe-based metal–organic framework as target recognition elements and enzyme mimics, respectively. The method demonstrated a broad detection range from 0.5 to 50 ng/mL, with a low detection limit of 0.03 ng mL−1, and exhibited a good recovery (91.0–102%) for bPAG9-spiked serum samples. Additionally, the aptasensor was successfully applied to detecting the pregnancy-specific biomarker bPAGs in serum samples.
期刊介绍：
As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.
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