The production and characterization of a monoclonal antibody (MAb AB10) against GA 3 glucoside as well as GA 3 is described. MAb AB10 was derived from an immunogen in which human serum albumin (HSA) was linked to G...The production and characterization of a monoclonal antibody (MAb AB10) against GA 3 glucoside as well as GA 3 is described. MAb AB10 was derived from an immunogen in which human serum albumin (HSA) was linked to GA 3 at carbon 3. This antibody showed high affinity for GA 3 glucoside as well as for 13 hydroxy gibberellins (GA 1, GA 3, GA 5, etc). The affinity of MAb AB10 for 13 hydroxy GAs was significantly reduced by methylation of the 7 oic acid but not by glycosylation of 3 hydroxyl group. Based on this antibody, both of competitive enzyme linked immunosorbent assays (ELISAs) for GA 3 glucoside and for GA 3 were developed. These two ELISAs displayed linear detection ranges from 0 2 pmol to 20 pmol. Using these assays, the fluctuation of GA 3 like and GA 3 glucoside like substances in the leaves of Rumex japonicus was investigated. The results indicated that the glycosylation of free GAs was connected with leaf senescence and that the function of 6 benzyl amino purine in retarding the leaf senescence was probably related to delaying the process of glycosylation of free GAs.展开更多
[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzym...[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in P.vannamei.[Results]The standard curve range of the kit was 0-8.1μg/L;the detection limit for P.vannamei was 0.912μg/kg;the recovery was 80.6%-103.5%;and the relative standard deviation range within batches was 5.3%-10.1%,and the relative standard deviation range between batches was 6.7%-8.1%.The specificity of the pentachloronitrobenzene monoclonal antibody was relatively good,and the cross-reaction rates with pentachlorophenol,hexachlorobenzene,tetrachlorophthalide,and chlorothalonil were low,all of which did not exceed 30%.The ELISA kit could be stored at 4℃for 12 months,showing good stability.[Conclusions]The detection kit has low cost,short time and small deviation,and is an ideal preliminary screening method.展开更多
[Objective] This study was conducted to find out an approach for determining trimethoprim residues in water. [Method] Trimethoprim antigen was prepared through a series of reactions from trimethoprim hapten which was ...[Objective] This study was conducted to find out an approach for determining trimethoprim residues in water. [Method] Trimethoprim antigen was prepared through a series of reactions from trimethoprim hapten which was generated through the reaction between trimethoprim and maleic anhydride. And trimethoprim monoclonal antibodies were prepared by animal immune, and used to prepare ELISA kit to detect trimethoprim residues in water. Finally, the limit of detection （LED） of the ELISA kit was determined. [Result] The standard curve covered a concentration range of 0-80 μg/L. The LeD of trimethoprim in water using the ELISA kit was 2.34 μg/kg; the IC50 （half maximal inhibitory concentration） was 4.8 μg/L; the recovery rate of added trimethoprim standard ranged from 60.5% to 79.7%; within-and among-batches RSD was less than 10%. The trimethoprim monoclonal antibody was specific, as the cross-reactivity rate of trimethoprim antibody and diaveridine was less than 1%. The stability tests revealed that the ELISA kit was stable after being stored at 4 ℃ for 12 months. [Conclusion] The results will provide references for controlling the abuse of trimethoprim.展开更多
目的建立肠道病毒71型(enterovirus type 71,EV-A71)灭活疫苗(Vero细胞)抗原含量检测方法并对其进行方法学验证及初步应用。方法以抗EV-A71兔多克隆抗体为包被抗体,辣根过氧化物酶(horseradish peroxidase,HRP)标记的抗EV-A71鼠单克隆...目的建立肠道病毒71型(enterovirus type 71,EV-A71)灭活疫苗(Vero细胞)抗原含量检测方法并对其进行方法学验证及初步应用。方法以抗EV-A71兔多克隆抗体为包被抗体,辣根过氧化物酶(horseradish peroxidase,HRP)标记的抗EV-A71鼠单克隆抗体为检测抗体,建立双抗体夹心ELISA并验证其线性范围、专属性、准确度、精密度及耐用性等。通过检测EV-A71疫苗原液及研发生产工艺各中间制品的EV-A71抗原含量评价该方法的适用性。结果包被抗体与酶标抗体最佳稀释度分别为1∶5000和1∶10000。抗原为5~80 U/mL时,线性良好;与同为肠道病毒的其他抗原不存在交叉反应,专属性良好;对不同浓度抗原进行6次测定,其回收率在90%~100%,准确度良好;不同实验员对不同浓度抗原样品检测3次,CV均<11%,精密度良好;针对不同影响因素设计耐用性试验,回收率均在80%~120%,耐用性良好。该方法检测EV-A71疫苗原液和制备过程中的中间制品均具有良好的线性和平行性,表明该方法具有良好的适用性。结论建立了EV-A71疫苗(Vero细胞)抗原含量检测方法并进行了初步应用,为EV-A71疫苗和HFMD多价疫苗的研发及质量控制提供了可靠的检测方法。展开更多
文摘The production and characterization of a monoclonal antibody (MAb AB10) against GA 3 glucoside as well as GA 3 is described. MAb AB10 was derived from an immunogen in which human serum albumin (HSA) was linked to GA 3 at carbon 3. This antibody showed high affinity for GA 3 glucoside as well as for 13 hydroxy gibberellins (GA 1, GA 3, GA 5, etc). The affinity of MAb AB10 for 13 hydroxy GAs was significantly reduced by methylation of the 7 oic acid but not by glycosylation of 3 hydroxyl group. Based on this antibody, both of competitive enzyme linked immunosorbent assays (ELISAs) for GA 3 glucoside and for GA 3 were developed. These two ELISAs displayed linear detection ranges from 0 2 pmol to 20 pmol. Using these assays, the fluctuation of GA 3 like and GA 3 glucoside like substances in the leaves of Rumex japonicus was investigated. The results indicated that the glycosylation of free GAs was connected with leaf senescence and that the function of 6 benzyl amino purine in retarding the leaf senescence was probably related to delaying the process of glycosylation of free GAs.
基金Key R&D Program of Hebei Province:Special Project on Key Common Technologies for High-quality Agricultural Development(20327505D).
文摘[Objectives]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in Penaeus vannamei.[Methods]This study was conducted to develop an enzyme-linked immunoassay kit that can detect the residual amount of pentachloronitrobenzene in P.vannamei.[Results]The standard curve range of the kit was 0-8.1μg/L;the detection limit for P.vannamei was 0.912μg/kg;the recovery was 80.6%-103.5%;and the relative standard deviation range within batches was 5.3%-10.1%,and the relative standard deviation range between batches was 6.7%-8.1%.The specificity of the pentachloronitrobenzene monoclonal antibody was relatively good,and the cross-reaction rates with pentachlorophenol,hexachlorobenzene,tetrachlorophthalide,and chlorothalonil were low,all of which did not exceed 30%.The ELISA kit could be stored at 4℃for 12 months,showing good stability.[Conclusions]The detection kit has low cost,short time and small deviation,and is an ideal preliminary screening method.
基金Supported by Beijing Municipal Science and Technology Project(Z151100002115059)~~
文摘[Objective] This study was conducted to find out an approach for determining trimethoprim residues in water. [Method] Trimethoprim antigen was prepared through a series of reactions from trimethoprim hapten which was generated through the reaction between trimethoprim and maleic anhydride. And trimethoprim monoclonal antibodies were prepared by animal immune, and used to prepare ELISA kit to detect trimethoprim residues in water. Finally, the limit of detection （LED） of the ELISA kit was determined. [Result] The standard curve covered a concentration range of 0-80 μg/L. The LeD of trimethoprim in water using the ELISA kit was 2.34 μg/kg; the IC50 （half maximal inhibitory concentration） was 4.8 μg/L; the recovery rate of added trimethoprim standard ranged from 60.5% to 79.7%; within-and among-batches RSD was less than 10%. The trimethoprim monoclonal antibody was specific, as the cross-reactivity rate of trimethoprim antibody and diaveridine was less than 1%. The stability tests revealed that the ELISA kit was stable after being stored at 4 ℃ for 12 months. [Conclusion] The results will provide references for controlling the abuse of trimethoprim.
文摘目的建立肠道病毒71型(enterovirus type 71,EV-A71)灭活疫苗(Vero细胞)抗原含量检测方法并对其进行方法学验证及初步应用。方法以抗EV-A71兔多克隆抗体为包被抗体,辣根过氧化物酶(horseradish peroxidase,HRP)标记的抗EV-A71鼠单克隆抗体为检测抗体,建立双抗体夹心ELISA并验证其线性范围、专属性、准确度、精密度及耐用性等。通过检测EV-A71疫苗原液及研发生产工艺各中间制品的EV-A71抗原含量评价该方法的适用性。结果包被抗体与酶标抗体最佳稀释度分别为1∶5000和1∶10000。抗原为5~80 U/mL时,线性良好;与同为肠道病毒的其他抗原不存在交叉反应,专属性良好;对不同浓度抗原进行6次测定,其回收率在90%~100%,准确度良好;不同实验员对不同浓度抗原样品检测3次,CV均<11%,精密度良好;针对不同影响因素设计耐用性试验,回收率均在80%~120%,耐用性良好。该方法检测EV-A71疫苗原液和制备过程中的中间制品均具有良好的线性和平行性,表明该方法具有良好的适用性。结论建立了EV-A71疫苗(Vero细胞)抗原含量检测方法并进行了初步应用,为EV-A71疫苗和HFMD多价疫苗的研发及质量控制提供了可靠的检测方法。
