Radioimmunoimaging of colorectal cancer using (99m)Tc labeled monoclonal antibody.
ABSTRACT AIM:To determine whether Hb3 and its fragment F(ab')(2) have practical value in radioimmunoimaging of colorectal cancer.METHODS:Intact Hb3 was purified by hydroxylapatite chromatography.The fragment F(ab') (2) was prepared by cold digestion and purified as intact Hb3.Hb3 and its fragment F(ab') (2) were labeled with 99mTc by direct labeling method using SnCl(2) as reducing agent. The radioactive doses ranged from 15 to 40 mCi.The imaging was accomplished by single photon emission computered tomograph (SPECT) with imaging time ranging from 2.5 to 48 hours. In this study, 10 patients were selected. Among them, 7 were administered with intact Hb3, and 3 with F(ab') (2) fragment. All the patients were diagnosed as having colorectal adenocarcinoma.RESULTS:After purification, intact Hb3 and its fragment F(ab') (2) were fit for radioimmunoimaging. The percentage of labeling of (99m)Tc to Hb3 or F(ab') (2) was 80.6%-91.5%. Among the 10 patients, 3 of 7 patients administered with intact Hb3 had positive scans, the other 4 had negative scans, and 2 of 3 patients administered with F(ab') (2)had positive scans, the other 1 had negative scans.CONCLUSION:The results showed that both intact Hb3 and its F(ab') (2) have some practical value in radioimmunoimaging of colorectal cancer, and the effects of imaging with F(ab') (2) was better than that with intact Hb3.
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ABSTRACT: Monoclonal antibodies of IgG1 subclass raised against purified human prostate-specific acid phosphatase were subjected to different procedures to produce F(ab')2 fragments suitable for radioimaging of prostatic cancer, following derivatization and labeling with radionuclides. The main aim was to obtain highly purified fragments with preserved immunological activity. Optimized pepsin digestion led to the formation of mainly F(ab')2 and Fab fragments, and, following Sephacryl S-200 gel filtration, the yield of pure F(ab')2 fragments was 24 +/- 11% of the theoretical maximum. After papain digestion in the absence of thiols, no formation of Fab fragments was observed, and the F(ab')2 fragments formed could be efficiently separated from Fc fragments by chromatofocusing or ion exchange chromatography. The yield of F(ab')2 fragments from papain digestion was 50 +/- 5% of the theoretical maximum. Both the above procedures gave F(ab')2 fragments with immunoreactivity and affinity identical to those of the precursor IgG1, despite the fact that isoelectric focusing profiles of the two F(ab')2 preparations differed, suggesting different digestion sites.Journal of Immunological Methods 07/1988; 110(2):229-36. · 2.23 Impact Factor
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ABSTRACT: Background. Previous studies in the literature have suggested that radiolabeled F(ab′)2 fragments might be superior to whole immunoglobulin G (IgG) for imaging and therapy of cancer because of their greater penetration in tumors. To test this hypothesis, the authors compared tumor and normal tissue uptake along with plasma clearance of 125I-labeled monoclonal antibody (MoAb) IMMU-4 whole IgG with its 131I-labeled F(ab′)2 fragment.Methods. Five patients with either liver metastases from colorectal cancer (n = 4) or intact primary tumors (n = 1) received a combination of 125I-IMMU-4 IgG (2 mCi/1 mg) plus131I-IMMU-4 F(ab′)2 (10 mCi/1 mg) as a single 1-hour intravenous infusion on day 1. Serial blood samples were taken for up to 72 hours postinfusion to determine plasma clearance of each MoAb. On days 3–9, patients underwent exploratory laparotomy in which biopsies of tumor as well as normal tissues (liver, normal colon, lymph node, and blood) were obtained. Tissues were weighed and counted in a gamma counter, and the percent of injected dose per kilogram (%ID/kg) of each antibody, along with the radiolocalization index (RI), was computed (RI = %ID/kg tumor. %ID/kg normal tissue).Results. Tumor uptake of both antibodies (2.3 ± 0.53 %ID/kg) was significantly higher than that of normal tissues (0.56 ± 0.12; P < 0.001), except for blood (2.8 ± 0.83), resulting in an RI ≥ 3. There were no significant differences in uptake (%ID/g) between F(ab′)2 and IgG (F[ab′]2 = 2.0 ± 0.57; IgG = 2.6 ± 0.94). The mean ± SD of plasma T½ was slightly shorter for F(ab′)2 (28.8 ± 7.2 hours) than for IgG (45.9 ± 16.7; P = 0.08).Conclusion. In short, the biodistribution and pharmacokinetics of IMMU-4 F(ab′)2 were comparable to those of IMMU-4-IgG. Cancer 1994; 73:850–7.Cancer 06/2006; 73(S3):850 - 857. · 5.20 Impact Factor
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ABSTRACT: The direct labeling of antibodies and antibody fragments to form a highly stable bond between technetium and the sulfide groups of proteins is now well established. To optimize this reaction, the antibody protein must have sufficient reactive sulfides available to accept that technetium metal ions that are formed by the reduction of pertechnetate in the presence of a weak complexing agent. The reactive sulfide groups are provided by first reducing a small fraction of the disulfide bridges in the antibody protein or by starting with Fab' fragments, which already have reactive sulfide groups. When the antibody protein has been appropriately reduced, and the reactive sulfide groups protected by a metal ion with a lower binding affinity than technetium, such as tin or zinc, very high labeling yields of high-affinity-bonded 99mTc can be achieved. This can be accomplished without loss of immunoreactivity, measured as either affinity or immunoreactive fraction. Side reactions can produce radiochemical impurities such as low-affinity, bound 99mTc; 99mTc colloids; 99mTc peptides or antibody aggregates; or 99mTc-complexes. Also, pertechnetate ions may be an impurity if the sodium pertechnetate solution added to the reduced antibodies is not completely reduced. The specifics of minimizing these side reactions have not been extensively discussed in the prior literature; however, it is clear that appropriate reduction of the protein prior to labeling and complete removal of the reducing agent, particularly if it contains reactive sulfide groups or is toxic, are critical. One- or two-step 99mTc-labeling kits for preparing 99mTc-labeled antibody or antibody fragments are rapidly being introduced for use in clinical nuclear medicine studies. These direct labeling methods employ a common sequence of chemical reactions, although the reducing agents for both the antibody and the [99mTc]pertechnetate may vary. Different 99mTc transfer agents may be used, but all transfer agents have the common feature of quickly forming weak to moderately strong complexes with reduced technetium. Most use Sn(II) to reduce the pertechnetate, although other reducing agents can be used.International Journal of Radiation Applications and Instrumentation Part B Nuclear Medicine and Biology 02/1991; 18(7):667-76.