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
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).
