磁免疫结合单颗粒模式ICP-MS同时测定乳腺癌病人血清中的CEA与CA15-3  

Detection of Carcinoembryonic Antigen and Carbohydrate Antigen 15-3 in Human Breast Cancer Serum Using Single Particle Mode ICP-MS Coupled with Magnetic Immunoassay

作　　者：何永昱 曹玉嫔 覃东庙 邓必阳 HE Yong-yu;CAO Yu-pin;QIN Dong-miao;DENG Bi-yang(State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources,School of Chemistry and Pharmaceutical Sciences,Guangxi Normal University,Guilin 541004,China;College of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530200,China)

机构地区：[1]广西师范大学化学与药学学院省部共建药用资源化学与药物分子工程国家重点实验室,广西桂林541004 [2]广西中医药大学药学院,广西南宁530200

出　　处：《分析测试学报》2021年第6期965-972,共8页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(21765004)。

摘　　要：该文合成了氨基修饰的磁性纳米颗粒并将其与癌胚抗原(CEA)一抗和糖类抗原15-3(CA15-3)一抗偶联用提取靶标生物标志物,羧基功能化上转换纳米颗粒(UCNPs)和氨基功能化二氧化铈-二氧化硅纳米颗粒(CeO2-SiO2NPs)用于标记抗体,由于174Yb和140Ce的瞬态信号频率与UCNPs和CeO2-SiO2NPs浓度直接相关,可利用单颗粒ICP-MS产生的瞬态信号频率定量分析CEA和CA15-3,基于此建立了同时测定乳腺癌病人血清中CEA和CA15-3的单颗粒ICP-MS磁免疫分析方法。结果显示:在优化条件下,CEA和CA15-3分别在0.02~100 ng·mL^(-1)和0.05~50 U·mL^(-1)浓度范围内线性良好,相关系数(r2)分别为0.9940和0.9870;以3倍信噪比(S/N=3)计算得CEA和CA15-3的检出限(LOD)分别为0.0067 ng·mL^(-1)和0.0167 U·mL^(-1)。采用该方法在3名乳腺癌病人血清中均检出CEA和CA15-3,加标回收率分别为95.2%~98.9%和95.5%~97.2%,相对标准偏差(RSD)分别为3.0%~4.6%和3.4%~4.3%。实验结果与临床使用的化学发光免疫方法结果一致,方法可用于乳腺癌病人血清样品中CEA和CA15-3含量的测定。Carcinoembryonic antigen(CEA)is a kind of glycoprotein that has been extensively investigated as a tumor marker for clinical diagnosis,which is routinely used as part of an individual's annual medical checkup in China.It is present in about 95%of all colon tumors and 50%of breast tumors and is also associated with ovarian carcinoma,lung cancer and other cancers.CEA is a preferred tumor marker to help predict outlook in patients with colorectal cancer.Breast cancer is the most widespread life-threatening cancer among women worldwide.It is a complex and heterogeneous disease composed of several subtypes with different morphological and clinical implications.One of the most important biomarkers for breast cancer is the carbohydrate antigen 15-3(CA15-3).The concentration of CA15-3 in blood serum could provide information from monitoring of therapy,which could also predict breast cancer recurrence after curative surgery.Hence,it is important to develop a highly sensitive analytical method for accurate and selective detection of CEA and CA15-3.Compared to one element tagging,the immunoassay of multi-elements can improve assay efficiency and reduce sample consumption.In this study,several immunoassay procedure parameters were examined,including effects of dwell time,concentrations of CeO2-SiO2NPs and UCNPs on the number of detection events.A new method for simultaneous determination of CEA and CA15-3 was developed using single particle inductively coupled plasma mass spectrometry(ICP-MS)coupled with magnetic immunoassay.Meanwhile,amino-modified magnetic nanoparticles were synthesized and conjugated with primary CEA antibody and primary CA15-3 antibody to extract the target biomarkers.The suspension solutions were separated with a permanent magnet.Single particles were directly introduced into ICP-MS with a custom,gas pressure-assisted sample introduction system.Carboxyl-functionalized up-conversion nanoparticles(UCNPs)and amino-functionalized cerium dioxide-silicon dioxide nanoparticles(CeO2-SiO2NPs)were employed to tag

关 键 词：电感耦合等离子体质谱(ICP-MS) 纳米粒子 癌胚抗原(CEA) 糖类抗原15-3(CA15-3) 血清 

分 类 号：O657.63[理学—分析化学]