AIM To directly radiolabel an anti-hepatomamAb fragment HAb18 F（ab’）<sub>2</sub> with <sup>99m</sup>Tc bystannous-reduced method,and assess thestability,biodistribution and radioimmun-oimag...AIM To directly radiolabel an anti-hepatomamAb fragment HAb18 F（ab’）<sub>2</sub> with <sup>99m</sup>Tc bystannous-reduced method,and assess thestability,biodistribution and radioimmun-oimaging（RⅡ）.METHODS Immunoreactive fraction wasdetermined according to Lindmo’s method.Ellman’s reagent was used to determine thenumber of thiols in the reduced F（ab’）<sub>2</sub>.Labelingefficiency and homogeneity were measured bypaper chromatography,sodium dodecylsulphatepolyacrylamide gel electrophoresis（SDS-PAGE）and autoradiography.Challenge assay involvedthe incubation of aliquots of labeled antibody inethylenediaminetetraacetate（ EDTA ）and L-cysteine（L-cys）solutions with different molarratio at 37℃ for 1h,respectively.Investigationsin vivo utilized nude mice bearing humanhepatocellular carcinoma（HHCC）xenograftswith gamma camera imaging and tissuebiodistribution studies at regular intervals.RESULTS The labeling procedure was finishedwithin 1.5 h compared with the'pretinning'method which would take at least 21h.In vitrostudies demonstrated that the radiolabeled mAbfragment was homogeneous and retained itsimmunoreactivity.Challenge studies indicatedthat <sup>99m</sup>Tc-labeled HAb18 F（ab’）<sub>2</sub> in EDTA is morestable than in L-cys.Imaging and biodistribution showed a significant tumor uptake at 24 h post-injection of <sup>99m</sup>Tc-labeled HAb18 F（ab’）<sub>2</sub>.Theblood,kidney,liver and tumor uptakes at 24hwere 0.56±0.09,56.45±11.36,1.43±0.27 and6.57±3.01（%ID/g）,respectively.CONCLUSION <sup>99m</sup>Tc-HAb18 F（ab’）<sub>2</sub> conjugateprepared by this direct method appears to be aneffective way to detect hepatoma in nude micemodel.展开更多
目的制备膀胱癌特异性核基蛋白BLCA-4单克隆抗体。方法根据已确定的膀胱特癌异性核基蛋白BLCA-4的特异性氨基酸片段序列：羧基端－EISQLNAGAC－氨基端，进行多肽合成，将多肽用Imjeet Immtmogen EDC Conjugation Kit耦联KLH作为抗原，...目的制备膀胱癌特异性核基蛋白BLCA-4单克隆抗体。方法根据已确定的膀胱特癌异性核基蛋白BLCA-4的特异性氨基酸片段序列：羧基端－EISQLNAGAC－氨基端，进行多肽合成，将多肽用Imjeet Immtmogen EDC Conjugation Kit耦联KLH作为抗原，用杂交瘤细胞制备BLCA-4单克隆抗体结果得到纯化抗BLCA-4单克隆抗体3F10及2F11。3F10表达于膀胱尿路上皮癌及癌旁尿路上皮细胞核内，正常膀胱尿路上皮无表达。结论制备的膀胱癌特异性核基蛋白BLCA-4单克隆抗体初步结果显示有较高的敏感性和特异性。展开更多
基金National Natural Science Foundation of China,No.39700175
文摘AIM To directly radiolabel an anti-hepatomamAb fragment HAb18 F（ab’）<sub>2</sub> with <sup>99m</sup>Tc bystannous-reduced method,and assess thestability,biodistribution and radioimmun-oimaging（RⅡ）.METHODS Immunoreactive fraction wasdetermined according to Lindmo’s method.Ellman’s reagent was used to determine thenumber of thiols in the reduced F（ab’）<sub>2</sub>.Labelingefficiency and homogeneity were measured bypaper chromatography,sodium dodecylsulphatepolyacrylamide gel electrophoresis（SDS-PAGE）and autoradiography.Challenge assay involvedthe incubation of aliquots of labeled antibody inethylenediaminetetraacetate（ EDTA ）and L-cysteine（L-cys）solutions with different molarratio at 37℃ for 1h,respectively.Investigationsin vivo utilized nude mice bearing humanhepatocellular carcinoma（HHCC）xenograftswith gamma camera imaging and tissuebiodistribution studies at regular intervals.RESULTS The labeling procedure was finishedwithin 1.5 h compared with the'pretinning'method which would take at least 21h.In vitrostudies demonstrated that the radiolabeled mAbfragment was homogeneous and retained itsimmunoreactivity.Challenge studies indicatedthat <sup>99m</sup>Tc-labeled HAb18 F（ab’）<sub>2</sub> in EDTA is morestable than in L-cys.Imaging and biodistribution showed a significant tumor uptake at 24 h post-injection of <sup>99m</sup>Tc-labeled HAb18 F（ab’）<sub>2</sub>.Theblood,kidney,liver and tumor uptakes at 24hwere 0.56±0.09,56.45±11.36,1.43±0.27 and6.57±3.01（%ID/g）,respectively.CONCLUSION <sup>99m</sup>Tc-HAb18 F（ab’）<sub>2</sub> conjugateprepared by this direct method appears to be aneffective way to detect hepatoma in nude micemodel.
文摘目的制备膀胱癌特异性核基蛋白BLCA-4单克隆抗体。方法根据已确定的膀胱特癌异性核基蛋白BLCA-4的特异性氨基酸片段序列：羧基端－EISQLNAGAC－氨基端，进行多肽合成，将多肽用Imjeet Immtmogen EDC Conjugation Kit耦联KLH作为抗原，用杂交瘤细胞制备BLCA-4单克隆抗体结果得到纯化抗BLCA-4单克隆抗体3F10及2F11。3F10表达于膀胱尿路上皮癌及癌旁尿路上皮细胞核内，正常膀胱尿路上皮无表达。结论制备的膀胱癌特异性核基蛋白BLCA-4单克隆抗体初步结果显示有较高的敏感性和特异性。
