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AIM To directly radiolabel an anti-hepatomamAb fragment HAb18 F(ab’)_2 with ~(99m)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’)_2.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 ~(99m)Tc-labeled HAb18 F(ab’)_2 in EDTA is morestable than in L-cys.Imaging and biodistribution showed a significant tumor uptake at 24 h post-injection of ~(99m)Tc-labeled HAb18 F(ab’)_2.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 ~(99m)Tc-HAb18 F(ab’)_2 conjugateprepared by this direct method appears to be aneffective way to detect hepatoma in nude micemodel.
AIM To direct radiolabel an anti-hepatomamAb fragment HAb18 F(ab’)_2 with ~(99m)Tc bystannous-reduced method, and assess thestability, biodistribution and radioimmun-oimaging(RII).METHODS Immunoreactive fraction wasdetermined according to Lindmo’s method.Ellman’s The reagent was used to determine the number of thiols in reduced F(ab’)_2.Labelingefficiency and homogeneity measured measured by paper chromatography, sodium dodecylsulphatepolyacrylamide gel electrophoresis (SDS-PAGE) and autoradiography.Challenge assay involved the incubation of aliquots of labeled antibody in ethylenediaminetetraacetate (EDTA )and L-cysteine(L-cys)solutions with Different molarratio at 37°C for 1h,respectively.Investigationsin vivoscope 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 vitrost Udies demonstrated that the radiolabeled mAbfragment was homogeneous and maintained itsimmunoreactivity.Challenge studies indicatedthat ~(99m)Tc-labeled HAb18 F(ab’)_2 in EDTA is morestable than in L-cys.Imaging and biodistribution showed a significant tumor uptake at 24 h Post-injection of ~(99m)Tc-labeled HAb18 F(ab’)_2.Theblood,kidney,liver and tumor uptakes at 24hwere 0.56±0.09,56.45±11.36,1.43±0.27 and 6.57±3.01(%ID/g ), respectively.CONCLUSION ~(99m)Tc-HAb18 F(ab’)_2 conjugateprepared by this direct method appears to be an effective way to detect hepatoma in nude micemodel.