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In order to construct a single chain fragment variable (ScFv) phage display library against ovarian tumor, by using RT-PCR, the human heavy chain variable region genes (VH) and light chain variable region genes (VL) were amplified from lymphocytes of ovarian tumor patients and subsequently assembled into ScFv genes by SOE. The resulting ScFv genes were electrotransformed into E.coli TG1 and amplified with the co-infection of helper phage M13KO7 to obtain phage display library. The capacity and titer of the resulting library were detected. The phage antibody library with a capacity of approximately 3 × 109 cfu/μg was obtained. After amplification with helper phage, the titer of antibody library reached 5 × 1012 cfu/mL. Human ScFv library against ovarian tumor was constructed successfully, which laid a foundation for the screening of ovarian tumor specific ScFv for the radioimmunoimaging diagnosis of ovarian tumor.
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Objective To study the distribution and tumor targeting property of anti-human prostate specific membrane antigen(PSMA) monoclonal antibody Ed-5 labeled with ~ 131 I in nude mice bearing human prostate cancer. Methods The nude mice xenografted models of human prostate cancer were established. Anti-human PSMA monoclonal antibody Ed-5 was labeled with ~ 131 I. ~ 131 I-Ed-5 was injected through the tail vein of nude mice bearing human prostate cancer. r-ray scintigraphy was performed at 24,48,96 and 144 hours after injection. And then the mouse were killed, the radioactivity ratio of tumor and non-tumor (T/NT), percentage of injected dosage of every gram tissue (%ID/g) were measured, and the bio-distribution of the labeled antibody was determined. Results The tumor imaging was clearly visualized at 96 hours after ~ 131 I-Ed-5 injection, and the tumor brim was clearer along with time prolonging. Radioactivity assembled in the tumors 24 to 144 hours after ~ 131 I-Ed-5 injection. The tumor's %ID/g at 96 hours after injection was 32.30, the radioactivity ratio of tumor and various tissues reached a maximum, and that of tumor/muscle was 7.08. Conclusion Ed-5 exhibited excellent immunoreactivity and tumor targeting property, and had application potential in targeting diagnosis and therapy of prostate cancer.
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Objective: To evaluate the targeting activity in the animal model with human hepatoma, the 131I-human antiHBsAg Fab radioimmunoimaging was explored. Methods: Radioimmunoimagings were taken on different intervals after injection of 131 I-human anti-HBsAg Fab to the nude mice and tissue distribution was measured. The human anti-HBsAg Fab was compared with the murine monoclonal antibodies. Results: The experimental group developed tumor positive images after 3 days of radio-labeled monoclonal antibodies injection, and the peak accumulation of radio-activity on the 5th day.Statistics indicated the tumor/liver ratio of the human anti-HBsAg Fab, murine monoclonal antibodies and the control groups were 5.4,4.0 and 0.9 respectively on the 7th day. Conclusions: Our results suggest that the 131 I-human anti-HB-sAg Fab has a considerable targeting activity, and provide an evidence that it can be used as a novel humanized carrer for targeting therapy of hepatoma.
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Objective:To label monoclonal anti-GPⅢa antibody (SZ21) with 99 Tcm and evaluate the value of 99 Tcm-SZ-21 for detecting thrombi in vivo.Methods:SZ-21 was modified with 2-iminotholane and labeled with 99 Tcm-GH and radiochemical purity was determined by ITLC-SG.99Tcm-SZ-21 was injected via ear edge vein in 5 rabbits in which thrombi were induced in the right femoral arteries.As control,99 Tcm-GH was administered in 1 rabbit.Static images were acquired and irregularly shaped ROIs were drawn on the images to calculate the ratios of T/M and T/B.Vein blood was drawn at 2 min,5 min,10 min,30 min and 1 h-3 h after injection in 2 rabbits so as to observe the blood clearance of 99 Tcm-SZ-21.Rabbits were sacrificed after 3 h of imaging.The vessels including clots were harvested and imaged.Cardiac muscle,liver,spleen,lung,kidney,etc.were excised and weighed.Radioactivity counts were measured to calculate % ID/g.Results:The radiochemical purity of 99 Tcm-SZ21 was beyond 90% and stable in vitro.Thrombi could be visualized at 30 min after injection,and at 2 h image of thrombi was clearly visualized,T/M (2.55?0.72) and T/B (1.94?0.51) ratios were high.In vitro imaging showed that T/B was 4.43?1.5.Conclusion:99Tcm-SZ-21 could be a potential agent for imaging diagnosis of thrombotic disease.
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Biodistribution and radioimmunoimaging of monoclonal antibody HB8759 agaist human melanoma in tumor-bearing nudemice were investigated.The results showed that mean tumor: nontumor(5 main organs)ratios were 6.6 and 9.6 at 48 and 72hr after injection of the ~(131)Ⅰ—McAb,respectively,and tumor was clearly visualized under ECF at the same time.It is suggested that McAb HB8759 is a potential biologic product for diagnosis of metastic melanoma and intraocularmelanoma and clinical treatment of melanoma.