Nutriology relies on advanced analytical tools to study the molecular compositions of food and provide key information on sample quality/safety. Small nutrients detection is challenging due to the high diversity and b...Nutriology relies on advanced analytical tools to study the molecular compositions of food and provide key information on sample quality/safety. Small nutrients detection is challenging due to the high diversity and broad dynamic range of molecules in food samples, and a further issue is to track low abundance toxins. Herein, we developed a novel plasmonic matrix-assisted laser desorption/ionization mass spectrometry(MALDI MS)approach to detect small nutrients and toxins in complex biological emulsion samples. Silver nanoshells(SiO_2@-Ag) with optimized structures were used as matrices andachieved direct analysis of ~ 6 n L of human breast milk without any enrichment or separation. We performed identification and quantitation of small nutrients and toxins with limit-of-detection down to 0.4 pmol(for melamine) and reaction time shortened to minutes, which is superior to the conventional biochemical method currently in use. The developed approach contributes to the near-future application of MALDI MS in a broad field and personalized design of plasmonic materials for real-case bio-analysis.展开更多
目的探讨基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)快速检测肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶的临床价值。方法收集2022年1月至2023年10月徐州医科大学附属邳州市人民医院分离的肺炎克雷伯菌60株和铜绿假单胞菌80株,其中...目的探讨基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)快速检测肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶的临床价值。方法收集2022年1月至2023年10月徐州医科大学附属邳州市人民医院分离的肺炎克雷伯菌60株和铜绿假单胞菌80株,其中碳青霉烯类耐药肺炎克雷伯菌(CRKP)30株,碳青霉烯类敏感肺炎克雷伯菌(CSKP)30株,碳青霉烯类耐药铜绿假单胞菌(CRPA)50株,碳青霉烯类敏感铜绿假单胞菌(CSPA)30株。分别采用改良碳青霉烯灭活试验(mCIM)、胶体金免疫层析法和Autof MS 1000质谱鉴定系统进行检测,评估Autof MS 1000质谱鉴定系统检测肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶的能力。结果Autof MS 1000质谱鉴定系统检测结果与mCIM和胶体金免疫层析法结果完全一致,30株CRKP中,有28株检测到碳青霉烯酶,2株阴性;50株CRPA中,有15株检测到碳青霉烯酶,35株阴性;30株CSKP和30株CSPA均为阴性。3种方法检测碳青霉烯酶的结果符合率为100%。结论Autof MS 1000质谱鉴定系统检测碳青霉烯酶的结果与mCIM法和胶体金免疫层析法一致,且既有mCIM法低成本的特点,又有胶体金免疫层析技术速度快、准确率高的优点,可用于临床肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶菌株的快速鉴定。展开更多
利用DNA重组技术在柞蚕蛹中表达重组人促红细胞生成素(Recombinant human erythropoietin,rhEPO),经过亲和层析纯化后,用SDS-PAGE分离纯化各组分,并通过电喷雾电离串联质谱技术(ESI-MS/MS)检测其糖基化修饰.结果显示,在柞蚕蛹中表达的rh...利用DNA重组技术在柞蚕蛹中表达重组人促红细胞生成素(Recombinant human erythropoietin,rhEPO),经过亲和层析纯化后,用SDS-PAGE分离纯化各组分,并通过电喷雾电离串联质谱技术(ESI-MS/MS)检测其糖基化修饰.结果显示,在柞蚕蛹中表达的rhEPO比活性约为1190 U/μg,其糖基化修饰位点与人体表达的EPO一致,有3个N-糖基化位点和1个O-糖基化位点.凝集素杂交实验结合质谱结果表明,柞蚕蛹表达的rhEPO的糖链中缺乏唾液酸修饰,而缺少唾液酸修饰的EPO通过鼻腔给药后在多种神经系统疾病的治疗中发挥着重要的作用.所得结果为进一步研究外源蛋白在柞蚕蛹-柞蚕核型多角体病毒(Anthraea pernyi nucleopolyhedrorirus,ApNPV)宿主载体表达系统表达后的糖基化与生物活性提供了依据.展开更多
CZE ESI MS is one of the most sensitive detection method and one of the potential method for protomics analysis. In this work, peptide mixture was separated and detected by using CZE ESI MS, and the influence of pH va...CZE ESI MS is one of the most sensitive detection method and one of the potential method for protomics analysis. In this work, peptide mixture was separated and detected by using CZE ESI MS, and the influence of pH value of buffer and composition of sheath liquid on CE separation and MS detection was investigated. The method allows the detection limits up to 4 2—33 pg. The result of this method is better than that of CE UV for the peptide analysis.展开更多
基金the financial support from Project 81771983, 81750110544, 81750410695, 81650110523, and 81471096 (to LXQ) by National Natural Science Foundation of China (NSFC)Project 16441909300 by Shanghai Science and Technology Commission+2 种基金Project 2017YFC0909000 by Ministry of Science and Technology of Chinasponsored by the Program for Professor of Special Appointment (Eastern Scholar) at Shanghai Institutions of Higher Learning (TP2015015)supported by 14DZ2272400 Shanghai Key Laboratory of Pediatric Gastroenterology and Nutrition (to WC)
文摘Nutriology relies on advanced analytical tools to study the molecular compositions of food and provide key information on sample quality/safety. Small nutrients detection is challenging due to the high diversity and broad dynamic range of molecules in food samples, and a further issue is to track low abundance toxins. Herein, we developed a novel plasmonic matrix-assisted laser desorption/ionization mass spectrometry(MALDI MS)approach to detect small nutrients and toxins in complex biological emulsion samples. Silver nanoshells(SiO_2@-Ag) with optimized structures were used as matrices andachieved direct analysis of ~ 6 n L of human breast milk without any enrichment or separation. We performed identification and quantitation of small nutrients and toxins with limit-of-detection down to 0.4 pmol(for melamine) and reaction time shortened to minutes, which is superior to the conventional biochemical method currently in use. The developed approach contributes to the near-future application of MALDI MS in a broad field and personalized design of plasmonic materials for real-case bio-analysis.
文摘目的探讨基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)快速检测肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶的临床价值。方法收集2022年1月至2023年10月徐州医科大学附属邳州市人民医院分离的肺炎克雷伯菌60株和铜绿假单胞菌80株,其中碳青霉烯类耐药肺炎克雷伯菌(CRKP)30株,碳青霉烯类敏感肺炎克雷伯菌(CSKP)30株,碳青霉烯类耐药铜绿假单胞菌(CRPA)50株,碳青霉烯类敏感铜绿假单胞菌(CSPA)30株。分别采用改良碳青霉烯灭活试验(mCIM)、胶体金免疫层析法和Autof MS 1000质谱鉴定系统进行检测,评估Autof MS 1000质谱鉴定系统检测肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶的能力。结果Autof MS 1000质谱鉴定系统检测结果与mCIM和胶体金免疫层析法结果完全一致,30株CRKP中,有28株检测到碳青霉烯酶,2株阴性;50株CRPA中,有15株检测到碳青霉烯酶,35株阴性;30株CSKP和30株CSPA均为阴性。3种方法检测碳青霉烯酶的结果符合率为100%。结论Autof MS 1000质谱鉴定系统检测碳青霉烯酶的结果与mCIM法和胶体金免疫层析法一致,且既有mCIM法低成本的特点,又有胶体金免疫层析技术速度快、准确率高的优点,可用于临床肺炎克雷伯菌和铜绿假单胞菌产碳青霉烯酶菌株的快速鉴定。
文摘CZE ESI MS is one of the most sensitive detection method and one of the potential method for protomics analysis. In this work, peptide mixture was separated and detected by using CZE ESI MS, and the influence of pH value of buffer and composition of sheath liquid on CE separation and MS detection was investigated. The method allows the detection limits up to 4 2—33 pg. The result of this method is better than that of CE UV for the peptide analysis.
