表面等离子体共振技术快速筛检人血清蛋白质表达的方法应用
Application of Surface Plasmon Resonance Technology in Screening Protein Expression of Human Serum Samples
-
摘要:
目的 建立用表面等离子体共振(surface plasmon resonance, SPR)技术快速筛检大样本人群血清蛋白质表达的方法,并用该法检测塑料垃圾拆解回收工人血清蛋白质是否存在异常表达。 方法 选取国内某地区从事塑料垃圾拆解回收的208名职业工人为暴露人群,生活水平和生活习惯相近且无已知污染源的197名农业人口为对照人群。用SPR技术检测研究对象血清中HSP90、HSP70、HSP60、CYP1A1和XPA蛋白质含量。 结果 人血清HSP90、HSP70、HSP60、CYP1A1和XPA蛋白质均可检出;用磷酸盐蛋白缓冲液(phosphate buffer solution, PBST)以1:100体积比稀释血清后即可进行检测,检测流速为25μL/min。暴露人群的血清中HSP60、HSP70、CYP1A1蛋白质含量均高于对照人群(P <0.05)。 结论 SPR技术可用于检测大样本人群血清蛋白质的表达,且具有用样量少、高通量、快速、灵敏的优势;从事塑料垃圾拆解回收的工人血清HSP60、HSP70、CYP1A1蛋白质表达量较高,可能与其职业环境中污染物暴露有关。 Abstract:Objective To develop a quick method of screening the protein expressions in human serum samples, and to apply the method for detecting serum protein expressions among workers who engaged in plastic recycling. Methods Serum samples were collected from 208 workers who engaged in plastic recycling (as the exposure group)and 197 local farmers without exposure to known pollutants in their living environment (as the control group). Serum protein expressions of HSP90, HSP70, HSP60, CYP1A1 and XPA were detected using Surface Plasmon Resonance (SPR). Results Levels of HSP90, HSP70, HSP60, CYP1A1 and XPA proteins were detected. For serum samples, the optimal dilution ratio was 1:100 (Serum:PBST)and the running flow rate was 25 μL/min. The results showed that protein expressions of HSP70, HSP60 and CYP1A1 in exposure group were significantly increased compared with the control group (P < 0.05). Conclusion For large samples, the SPR has the advantages of high throughput, sensitive and less blood amount requirement when protein expressions are to be detected. An over expressions of serum proteins of HSP60, HSP70 and CYP1A1 among workers may relate to their occupational exposures.
下载:
