| 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 |
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| 基金项目:国家自然科学基金资助项目(21765004) |
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| 中文摘要: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0.994 0和0.987 0；以3倍信噪比(S/N=3)计算得CEA和CA15-3的检出限(LOD)分别为0.006 7 ng·mL-1和0.016 7 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。 |
| 中文关键词: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 |
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| Detection of Carcinoembryonic Antigen and Carbohydrate Antigen 15-3 in Human Breast Cancer Serum Using Single Particle Mode ICP-MS Coupled with Magnetic Immunoassay |
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| Abstract: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 individuals 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 antibodies.The frequencies of transient signals of 174Yb and 140Ce were directly related to the concentrations of UCNPs and CeO2-SiO2NPs.CEA and CA15-3 could be quantified via the frequency of transient signal produced by single particle ICP-MS.Under the optimal conditions,the linear concentration ranges for CEA and CA15-3 were 0.02-100 ng·mL-1 and 0.05-50 U·mL-1,respectively.Their correlation coefficients were 0.994 0 and 0.987 0,respectively.The detection limits(LOD,S/N=3) of CEA and CA15-3 were 0.006 7 ng·mL-1 and 0.016 7 U·mL-1,respectively.The method was applied to the simultaneous detection of CEA and CA15-3 in human breast cancer serum with recoveries of 95.2%-98.9% and 95.5%-97.2%,and relative standard deviations(RSD) of 3.0%-4.6% and 3.4%-4.3%,respectively. |
| Key Words:inductively coupled plasma mass spectrometry(ICP-MS) nanoparticle carcinoembryonic antigen（CEA） carbohydrate antigen 15-3（CA15-3） serum |
| 引用本文：何永昱,曹玉嫔,覃东庙,邓必阳.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[J].分析测试学报,2021,40(6):965-972. |
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