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高等学校化学学报

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糖基印迹电化学传感器研制及乳腺癌PD-L1阳性外泌体快速检测

罗奎1,林珈希2,李建平1,2   

  1. 1. 桂林理工大学环境科学与工程学院 2. 桂林理工大学化学与生物工程学院
  • 收稿日期:2024-11-29 修回日期:2025-01-08 出版日期:2025-02-14 发布日期:2025-02-14
  • 通讯作者: 李建平 E-mail:likianping@263.net
  • 基金资助:
    国家自然科学基金项目（批准号：22264010）资助

Development of a glycosyl-imprinted sensor and rapid detection of PD-L1 positive exosomes in breast cancer

LUO Kui 1, LIN Jiaxi 2, LI Jianping 1, 2   

  1. 1 College of Environmental Science and Engineering, Guilin University of Technology 2 College of Chemistry and Bioengineering, Guilin University of Technology
  • Received:2024-11-29 Revised:2025-01-08 Online:2025-02-14 Published:2025-02-14
  • Contact: LI Jianping E-mail:likianping@263.net
  • Supported by:
    Supported by the National Natural Science Foundation of China (No. 22264010)

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摘要： 构建了一种快速识别检测乳腺癌PD-L1阳性外泌体的糖基印迹电化学传感器. 首先，以乳腺癌阳性外泌体糖蛋白PD-L1过表达的糖链Galβ1-4GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Gal为模板分子，3-氨基苯硼酸为功能单体，电聚合法制备了糖基印迹聚合物（GIP）. 洗脱去除模板分子后得到了可特异性识别PD-L1阳性外泌体的印迹膜. 利用铁氰化钾作为探针测量GIP电极的DPV电流值。用RIPA裂解外泌体扣除游离蛋白干扰，记录电流值变化ΔI。ΔI随着重吸附PD-L1阳性外泌体浓度的增大而逐渐降低，并与浓度的对数值呈线性正相关，检测范围为2.36 × 102 ~ 1.18 × 107 particles/mL，检出限为93 particles/mL. 该法已成功用于临床样本中乳腺癌PD-L1阳性外泌体的检测，其加标回收率为93. 82% ~ 105. 32%. 通过糖基化程度差异，该传感器可用于临床样本中乳腺癌的筛查。

关键词: 外泌体, 糖基印迹, 电化学传感器, PD-L1, 乳腺癌

Abstract: A glycosylated electrochemical sensor for rapid identification and detection of breast cancer PD-L1 positive exosomes was developed. First, glycosyl-imprinted polymers (GIP) were prepared by electropolymerization using glyco-Galβ1-4GlcNAcβ1-3Galβ1-4GlcNAcβ1-3Gal overexpressed by breast cancer positive exosome glycoprotein PD-L1 as template molecules and 3-aminophenylboronic acid as functional monomers. After elution and removal of template molecules, a imprinted membrane that can specifically recognize PD-L1 positive exosomes was obtained. Potassium ferricyanide was used as a probe to measure the DPV current value of the GIP electrode. Exosomes were cleaved with RIPA to deduct free protein interference, and the change in current value ΔI was recorded. ΔI decreased with the increase of the concentration of readsorbed PD-L1-positive exosomes, and was linearly positively related with the logarithmic value of the concentration. The detection range was 2.36 × 102 ~ 1.18 × 107 Particles/mL, and the detection limit was 93 particles/mL. The method has been successfully used to detect breast cancer PD-L1-positive exosomes in clinical samples, and its spiked recoveries were 93. 82% ~ 105. 32%. The sensor could be used to screen breast cancer in clinical samples by the difference of glycosylation degree.

Key words: Exosome, Glycosyl-imprinting; Electrochemical sensor, PD-L1, Breast cancer

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