| 倪林君,刘晓凤,刘维清,胡筱,余丽双.基于抗原-抗体免疫反应构建荧光传感器检测地龙多肽[J].井冈山大学自然版,2025,46(2):35-41 |
| 基于抗原-抗体免疫反应构建荧光传感器检测地龙多肽 |
| A FLUORESCENCE SENSOR BASED ON ANTIGEN-ANTIBODY IMMUNOREACTION WAS CONSTRUCTED FOR THE DETECTION OF EARTHWORM POLYPEPTIDES |
| 投稿时间：2024-06-22 修订日期：2024-10-16 |
| DOI：10.3969/j.issn.1674-8085.2025.02.005 |
| 中文关键词: 地龙 荧光免疫分析 多肽检测 |
| 英文关键词: earthworm fluorescent immunoassay polypeptide detection |
| 基金项目:国家自然科学基金项目(82074005)；福建省科技厅科技计划项目(2021J01925) |
| 作者 | 单位 | E-mail | | 倪林君 | 福建中医药大学药学院, 福建, 福州 350122 | | | 刘晓凤 | 福建中医药大学药学院, 福建, 福州 350122 | | | 刘维清 | 福建中医药大学药学院, 福建, 福州 350122 | | | 胡筱 | 福建中医药大学药学院, 福建, 福州 350122 | xiao_hu@fjtcm.edu.cn | | 余丽双 | 福建中医药大学药学院, 福建, 福州 350122 | yuls66@fjtcm.edu.cn |
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| 中文摘要: |
| 基于多肽及其多克隆抗体结合形成的复合物的荧光特性，首次建立了一种可以快速检测地龙多肽 TP-21的免疫荧光传感器。地龙多肽 TP-21 和多克隆抗体具有较弱荧光，而两者结合形成的复合物具有较强荧光，通过检测复合物荧光强度可推断 TP-21 多肽浓度。实验研究对激发波长、缓冲体系、多克隆抗体浓度等条件进行了优化，在最佳条件下，复合物与多克隆抗体的荧光强度差值具有良好线性关系，线性范围为 0.058 59~30.00 μg·mL-1，R2= 0.992 7，检测限为 49.22 ng·mL-1。该方法操作简便、快速、灵敏度高，对于 TP-21 多肽具有良好选择性。实现实际样品地龙蛋白粉中 TP-21 多肽的含量测定，其回收率在 85.40%~105.68%范围，实验表明测定结果准确可靠，具有良好分析应用前景。 |
| 英文摘要: |
| Based on the fluorescence characteristics of the complex formed by the binding of polypeptide and its polyclonal antibody, an immunofluorescent sensor for the rapid detection of earthworm polypeptide TP-21 was established for the first time. TP-21 polypeptide and polyclonal antibody had weak fluorescence, but their complex had strong fluorescence. The concentration of TP-21 polypeptide could be inferred by detecting the fluorescence intensity of the complex. The excitation wavelength, buffer system and polyclonal antibody concentration were optimized. Under the optimal conditions, the difference of fluorescence intensity between the complex and polyclonal antibody had a good linear relationship, the linear range was 0.058 59~30.00 μg·mL-1, R2= 0.992 7, and the detection limit was 49.22 ng·mL-1. The method is simple, rapid, sensitive and has good selectivity for TP-21 peptide. The content of TP-21 polypeptide in earthworm protein powder was determined. The recovery rates ranged from 85.40% to 105.68%. The results showed that the determination was accurate and reliable. |
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