Immunomagnetic microbeads for screening with flow cytometry and identification with nano-liquid chromatography mass spectrometry of ochratoxins in wheat and cereal.
Immunomagnetic microbeads for screening with flow cytometry and identification with nano-liquid chromatography mass spectrometry of ochratoxins in wheat and cereal.
复制标题
通过流式细胞术筛选的免疫磁性微粒，并用小麦和谷物中的ochratoxins snatetry和用纳米液色谱质谱法进行鉴定。
DOI：
10.1007/s00216-011-4974-7
复制
发表时间：
2011-07
影响因子：
4.3
通讯作者：
Nielen, Michel W. F.
中科院分区：
文献类型：
--
作者：
Aqai, Payam;Peters, Jeroen;Gerssen, Arjen;Haasnoot, Willem;Nielen, Michel W. F.
用于快速筛选食品污染物的多分析物结合分析需要对可疑样品中的化合物(S)进行质谱学鉴定。当在两种方法中使用相同的生物试剂时，可以获得最佳组合；此外，由于生物试剂的高成本，小型化是重要的。在一种新的直接抑制流式细胞仪免疫分析(FCIA)和免疫亲和分离的概念中，使用包被单抗(MAb)的超顺磁性微球，然后通过纳米液相色谱-四极杆飞行时间质谱仪(Nano-LC-Q-ToF-MS)进行鉴定。以小麦和谷物中的真菌毒素赭曲霉毒素A(OTA)及其交叉反应的霉菌毒素类似物为模型体系，经简单提取后，用FCIA和抗OTA单抗进行分析。OTA的检出限为0.15 ng/g，远低于欧盟规定的3 ng/g的最低最高水平。在免疫磁性分离方法中，使用了350倍以上的珠子来捕获样品提取物中的赭曲霉毒素。洗涤后，用小体积的酸化乙腈/水(2/8 v/v)将结合的赭曲霉毒素从单抗上解离，然后用纳米LC-Q-ToF-MS进行分离和鉴定。在筛选出的可疑天然污染样本中，OTA及其非氯化类似物赭曲霉毒素B被全扫描准确质谱仪成功识别，作为鉴定未知但交叉反应的新出现的真菌毒素的概念证明。由于小型化和生物亲和性分离，这一概念可能适用于使用其他更昂贵的生物反应剂，如运输蛋白和受体，以筛选和识别已知和未知(或隐蔽)出现的食品污染物。包被抗体的微珠
Multi-analyte binding assays for rapid screening of food contaminants require mass spectrometric identification of compound(s) in suspect samples. An optimal combination is obtained when the same bioreagents are used in both methods; moreover, miniaturisation is important because of the high costs of bioreagents. A concept is demonstrated using superparamagnetic microbeads coated with monoclonal antibodies (Mabs) in a novel direct inhibition flow cytometric immunoassay (FCIA) plus immunoaffinity isolation prior to identification by nano-liquid chromatography–quadrupole time-of-flight-mass spectrometry (nano-LC-Q-ToF-MS). As a model system, the mycotoxin ochratoxin A (OTA) and cross-reacting mycotoxin analogues were analysed in wheat and cereal samples, after a simple extraction, using the FCIA with anti-OTA Mabs. The limit of detection for OTA was 0.15 ng/g, which is far below the lowest maximum level of 3 ng/g established by the European Union. In the immunomagnetic isolation method, a 350-times-higher amount of beads was used to trap ochratoxins from sample extracts. Following a wash step, bound ochratoxins were dissociated from the Mabs using a small volume of acidified acetonitrile/water (2/8 v/v) prior to separation plus identification with nano-LC-Q-ToF-MS. In screened suspect naturally contaminated samples, OTA and its non-chlorinated analogue ochratoxin B were successfully identified by full scan accurate mass spectrometry as a proof of concept for identification of unknown but cross-reacting emerging mycotoxins. Due to the miniaturisation and bioaffinity isolation, this concept might be applicable for the use of other and more expensive bioreagents such as transport proteins and receptors for screening and identification of known and unknown (or masked) emerging food contaminants. Microbead coated with antibody
登录
查看更多内容
高通量悬浮芯片技术同时检测氯霉素、瘦肉精、17-β-雌二醇三种兽药
影响因子：
6.2
作者：
Liu, Nan;Su, Pu;Chao, Fuhuan
通讯作者：
Chao, Fuhuan
影响因子：
6.2
作者：
Hernández, MJ;García-Moreno, MV;Barroso, CG
通讯作者：
Barroso, CG
DOI：
10.1016/j.cccn.2005.06.023
发表时间：
2006-01
期刊：
Clinica chimica acta; international journal of clinical chemistry
影响因子：
--
作者：
Dunbar SA
通讯作者：
Dunbar SA
影响因子：
6.1
作者：
Barna-Vetro, I;Solti, L;Wolfling, A
通讯作者：
Wolfling, A
影响因子：
3.9
作者：
Kim, Sanghee;Pyo, Hyeon-Bong;Kim, Wan-Joong
通讯作者：
Kim, Wan-Joong