Dynamic tracking analysis of monoclonal antibodies(mAbs)biotransformation in vivo is crucial,as certain modifications could inactivate the protein and reduce drug efficacy.However,a particular challenge(i.e.immune rec...Dynamic tracking analysis of monoclonal antibodies(mAbs)biotransformation in vivo is crucial,as certain modifications could inactivate the protein and reduce drug efficacy.However,a particular challenge(i.e.immune recognition deficiencies)in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen.To address this limitation,a multi-epitope affinity technology utilizing the metal organic framework(MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region(CDR)mimotope peptide(HH24)and non-CDR mimotope aptamer(CH1S-6T)onto the surface of MOF@Au nanocomposite.Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology.Moreover,the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HCAsn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry(LC-QTOF-MS).Therefore,multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb.Particularly,the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids.Compared to the spiked phosphate buffer(PB)model,faster modification trends were monitored in the spiked serum and patients'sera due to the catalytic effect of plasma proteins and relevant proteases.Differences in peptide modification levels of trastuzumab in patients'sera were also monitored.In summary,the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb,contributing to improved understanding and paving the way for future research and clinical applications.展开更多
The overuse of antibiotics and antitumor drugs has resulted in more and more extensive pollution of water bodies with organic drugs,causing detrimental ecological effects,which have attracted attention towards effecti...The overuse of antibiotics and antitumor drugs has resulted in more and more extensive pollution of water bodies with organic drugs,causing detrimental ecological effects,which have attracted attention towards effective and sustainable methods for antibiotics and antitumor drug degradation.Here,the hybrid nanomaterial(g-C_(3)N_(4)@Fe/Pd)was synthesized and used to remove a kind of both an antibiotic and antitumor drug named mitoxantrone(MTX)with 92.0%removal efficiency,and the MTX removal capacity is 450 mg/g.After exposing to the hybrid material the MTX aqueous solution changed color from dark blue to lighter progressively,and LC-UV results of residual solutions showthat a newpeak at 3.0min(MTX:13.2min)after removal by g-C_(3)N_(4)@Fe/Pd appears,with the simultaneous detection of intermediate products indicating that g-C_(3)N_(4)@Fe/Pd indeed degrades MTX.Detailed mass spectrometric analysis suggests that the nuclear mass ratio decreased from 445.2(M+1H)to 126.0(M+1H),169.1(M+1H),239.2(M+1H),267.3(M+1H),285.2(M+1H),371.4(M+1H)and 415.2(M+1H),and the maximum proportion(5.63%)substance of all degradation products(126.0(M+1H))is 40-100 times less toxic than MTX.A mechanism for the removal and degradation of mitoxantrone was proposed.Besides,actual water experiments confirmed that the maximum removal capacity of MTX by g-C_(3)N_(4)@Fe/Pd is up to 492.4 mg/g(0.02 g/L,10 ppm).展开更多
【目的】玉米发酵生产燃料乙醇的副产物酒糟蛋白饲料(DDGS)常作为蛋白质替代饲料广泛用于动物生产中。随着DDGS产业链的扩大,其生产过程中所带来的抗生素的残留问题应该得到重视,但目前关于DDGS中抗生素的残留研究报道较少。对全国经营...【目的】玉米发酵生产燃料乙醇的副产物酒糟蛋白饲料(DDGS)常作为蛋白质替代饲料广泛用于动物生产中。随着DDGS产业链的扩大,其生产过程中所带来的抗生素的残留问题应该得到重视,但目前关于DDGS中抗生素的残留研究报道较少。对全国经营和使用环节的玉米DDGS进行抽样调查,对70种抗生素的残留含量进行分析测定,以了解目前中国饲料用DDGS中抗生素的残留情况。【方法】首先利用四级杆飞行时间质谱(quadrupole time-of-flight mass spectrometry,QTOF)建立了大环内酯类、氟喹诺酮类、磺胺类、喹恶啉类、β-内酰胺类、林可胺类、四环素类、酰胺醇类等共70种抗生素药物基于质量数、保留时间和特征碎片离子的数据库。在全国9个省、市、自治区抽取经营和使用环节的玉米DDGS样品74批次,筛查其中可能含有的抗生素药物。样品的前处理:取一定量的均质样品,加入乙腈和水的混合溶液进行提取,经涡旋、震荡、离心后,取上清液过NH2固相萃取柱,收集淋洗液N2气浓缩至一定体积,过0.22μm滤膜。样品的分析:首先利用QTOF建立的筛查列表对样品DDGS中可能残留的抗生素进行初步筛选,以确定残留抗生素的种类,而后采用液相色谱串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)对发现疑似含有抗生素残留的样品进行定量分析,以确定抗生素的残留量。【结果】欧盟指令2002/657/EC规定了在应用质谱仪对目标化合物进行确证时的准则。一个母离子为2分,一个子离子为2.5分。在本试验条件下所有抗生素的检测结果都能达到这个要求,符合对目标化合物的确证准则。经QTOF定性分析后,抽样的所有DDGS中有一批次样品中含有抗生素残留,种类为甲氧苄啶。样品母离子峰、二级特征离子碎片、出峰时间与标准品完全一致,样品和标准品所有提取碎片离子及离子丰度比一致。利用LC-MS/MS对残留抗生素进行定量分析后,确定其残留量为107.7 ng·g-1。甲氧苄啶在添加浓度为10—1 000ng·mL-1,平均添加回收率为75.2%—97.9%,线性相关系数大于0.99,日内标准偏差为2.8%—5%,日间标准偏差为8%—10%。【结论】利用QTOF建立DDGS中抗生素类药物的定性筛选方法可作为玉米DDGS中残留抗生素快速筛查方法。存在残留的甲氧苄啶抗生素,可能在玉米发酵生产DDGS前被添加,目的是抑制有害微生物的生长繁殖。抽样分析结果表明,目前中国玉米DDGS中抗生素残留的种类较少、残留量也较低,但DDGS中抗生素残留情况仍不容忽视,相关部门也未给出DDGS发酵过程中可使用的抗生素的种类范围、使用限量以及使用规范。此外,还应该研究直接采用QTOF进行定量分析的方法,以及研究包括维吉尼亚霉素在内的其他抗生素的QTOF分析方法。展开更多
建立血液中呋喃芬太尼的LC-QTOF-MS检验方法。采用Eclipse Plus C18(1.8μm,3.0X150mm)色谱柱,以水相A(0.1%(v/v)甲酸和5mM乙酸铵的水溶液)和有机相B(乙腈)为流动相梯度洗脱,质谱采用电喷雾正离子全扫描模式,对血液中的呋喃芬太尼分析...建立血液中呋喃芬太尼的LC-QTOF-MS检验方法。采用Eclipse Plus C18(1.8μm,3.0X150mm)色谱柱,以水相A(0.1%(v/v)甲酸和5mM乙酸铵的水溶液)和有机相B(乙腈)为流动相梯度洗脱,质谱采用电喷雾正离子全扫描模式,对血液中的呋喃芬太尼分析检验。方法回收率为76.44%~92%,线性回归方程为y=4355.5 x+2242.9,检出限为0.1 ng·mL^(-1),定量限为2 ng·mL^(-1)。本方法操作简单,可为定性定量分析检测血液中的呋喃芬太尼提供数据帮助。展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.:82373829,82273893,and 82173773)the Natural Science Foundation of Guangdong Province,China(Grant Nos.:2021A1515220099,and 2022A1515011576)+1 种基金the High-End Foreign Experts Project,China(Grant No.:G2021199005L)the Science and Technology Program of Guangdong Provincial Medical Products Administration,China(Grant Nos.:2023TDZ11,and 2022ZDB04).
文摘Dynamic tracking analysis of monoclonal antibodies(mAbs)biotransformation in vivo is crucial,as certain modifications could inactivate the protein and reduce drug efficacy.However,a particular challenge(i.e.immune recognition deficiencies)in biotransformation studies may arise when modifications occur at the paratope recognized by the antigen.To address this limitation,a multi-epitope affinity technology utilizing the metal organic framework(MOF)@Au@peptide@aptamer composite material was proposed and developed by simultaneously immobilizing complementarity determining region(CDR)mimotope peptide(HH24)and non-CDR mimotope aptamer(CH1S-6T)onto the surface of MOF@Au nanocomposite.Comparative studies demonstrated that MOF@Au@peptide@aptamer exhibited significantly enhanced enrichment capabilities for trastuzumab variants in comparison to mono-epitope affinity technology.Moreover,the higher deamidation ratio for LC-Asn-30 and isomerization ratio for HCAsn-55 can only be monitored by the novel bioanalytical platform based on MOF@Au@peptide@aptamer and liquid chromatography-quadrupole time of flight-mass spectrometry(LC-QTOF-MS).Therefore,multi-epitope affinity technology could effectively overcome the biases of traditional affinity materials for key sites modification analysis of mAb.Particularly,the novel bioanalytical platform can be successfully used for the tracking analysis of trastuzumab modifications in different biological fluids.Compared to the spiked phosphate buffer(PB)model,faster modification trends were monitored in the spiked serum and patients'sera due to the catalytic effect of plasma proteins and relevant proteases.Differences in peptide modification levels of trastuzumab in patients'sera were also monitored.In summary,the novel bioanalytical platform based on the multi-epitope affinity technology holds great potentials for in vivo biotransformation analysis of mAb,contributing to improved understanding and paving the way for future research and clinical applications.
基金Financial support from the National Natural Science Foundation of China (No.22176147)the National Science Fund for Excellent Young Scholars of China (No.21822607)+1 种基金the Fundamental Research Funds for Central Universities (No.22120230295)the State Key Laboratory for Pollution Control is acknowledged.
文摘The overuse of antibiotics and antitumor drugs has resulted in more and more extensive pollution of water bodies with organic drugs,causing detrimental ecological effects,which have attracted attention towards effective and sustainable methods for antibiotics and antitumor drug degradation.Here,the hybrid nanomaterial(g-C_(3)N_(4)@Fe/Pd)was synthesized and used to remove a kind of both an antibiotic and antitumor drug named mitoxantrone(MTX)with 92.0%removal efficiency,and the MTX removal capacity is 450 mg/g.After exposing to the hybrid material the MTX aqueous solution changed color from dark blue to lighter progressively,and LC-UV results of residual solutions showthat a newpeak at 3.0min(MTX:13.2min)after removal by g-C_(3)N_(4)@Fe/Pd appears,with the simultaneous detection of intermediate products indicating that g-C_(3)N_(4)@Fe/Pd indeed degrades MTX.Detailed mass spectrometric analysis suggests that the nuclear mass ratio decreased from 445.2(M+1H)to 126.0(M+1H),169.1(M+1H),239.2(M+1H),267.3(M+1H),285.2(M+1H),371.4(M+1H)and 415.2(M+1H),and the maximum proportion(5.63%)substance of all degradation products(126.0(M+1H))is 40-100 times less toxic than MTX.A mechanism for the removal and degradation of mitoxantrone was proposed.Besides,actual water experiments confirmed that the maximum removal capacity of MTX by g-C_(3)N_(4)@Fe/Pd is up to 492.4 mg/g(0.02 g/L,10 ppm).
文摘【目的】玉米发酵生产燃料乙醇的副产物酒糟蛋白饲料(DDGS)常作为蛋白质替代饲料广泛用于动物生产中。随着DDGS产业链的扩大,其生产过程中所带来的抗生素的残留问题应该得到重视,但目前关于DDGS中抗生素的残留研究报道较少。对全国经营和使用环节的玉米DDGS进行抽样调查,对70种抗生素的残留含量进行分析测定,以了解目前中国饲料用DDGS中抗生素的残留情况。【方法】首先利用四级杆飞行时间质谱(quadrupole time-of-flight mass spectrometry,QTOF)建立了大环内酯类、氟喹诺酮类、磺胺类、喹恶啉类、β-内酰胺类、林可胺类、四环素类、酰胺醇类等共70种抗生素药物基于质量数、保留时间和特征碎片离子的数据库。在全国9个省、市、自治区抽取经营和使用环节的玉米DDGS样品74批次,筛查其中可能含有的抗生素药物。样品的前处理:取一定量的均质样品,加入乙腈和水的混合溶液进行提取,经涡旋、震荡、离心后,取上清液过NH2固相萃取柱,收集淋洗液N2气浓缩至一定体积,过0.22μm滤膜。样品的分析:首先利用QTOF建立的筛查列表对样品DDGS中可能残留的抗生素进行初步筛选,以确定残留抗生素的种类,而后采用液相色谱串联质谱法(liquid chromatography-tandem mass spectrometry,LC-MS/MS)对发现疑似含有抗生素残留的样品进行定量分析,以确定抗生素的残留量。【结果】欧盟指令2002/657/EC规定了在应用质谱仪对目标化合物进行确证时的准则。一个母离子为2分,一个子离子为2.5分。在本试验条件下所有抗生素的检测结果都能达到这个要求,符合对目标化合物的确证准则。经QTOF定性分析后,抽样的所有DDGS中有一批次样品中含有抗生素残留,种类为甲氧苄啶。样品母离子峰、二级特征离子碎片、出峰时间与标准品完全一致,样品和标准品所有提取碎片离子及离子丰度比一致。利用LC-MS/MS对残留抗生素进行定量分析后,确定其残留量为107.7 ng·g-1。甲氧苄啶在添加浓度为10—1 000ng·mL-1,平均添加回收率为75.2%—97.9%,线性相关系数大于0.99,日内标准偏差为2.8%—5%,日间标准偏差为8%—10%。【结论】利用QTOF建立DDGS中抗生素类药物的定性筛选方法可作为玉米DDGS中残留抗生素快速筛查方法。存在残留的甲氧苄啶抗生素,可能在玉米发酵生产DDGS前被添加,目的是抑制有害微生物的生长繁殖。抽样分析结果表明,目前中国玉米DDGS中抗生素残留的种类较少、残留量也较低,但DDGS中抗生素残留情况仍不容忽视,相关部门也未给出DDGS发酵过程中可使用的抗生素的种类范围、使用限量以及使用规范。此外,还应该研究直接采用QTOF进行定量分析的方法,以及研究包括维吉尼亚霉素在内的其他抗生素的QTOF分析方法。
