Abstract
Objective To label the monoclonal antibody 4E5 with 131I and to evaluate the lethal effects of 131I-4E5 against B cell lymphoma.Methods 4E5 was radiolabeled with 131I using the Iodogen method at room temperature.The labeling efficiency and radiochemical purity was measured with trichloroacetic acid (TCA) precipitation,and the immune activity and stability of 131I-4E5 was analyzed.The lethal effects of 131I-4E5 and 4E5 against Raji cells were evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTT) assay.Analysis of variance and t-test were used for data analysis with SPSS 13.0.Results The labeling rate of 131I-4E5 was (78.3±2.4)％,and the radiochemical purity was (95.7±1.8)％.Its specific activity and radioactive concentration were 0.58 MBq/μg and 3.90 × 1010 Bq/L,respectively.The radiochemical purity of 131I-4E5 mixed with serum and PBS was over 90％ after three days.The maximum specific binding efficiency of 131I-4E5 with Raji cells was (36.06±2.63)％.131I-4E5 exhibited a dose-dependent cytotoxicity against Raji cells.The lethal effect of the high dose group was significantly stronger than that of the low dose group.When the radioactive concentrations were 1.48 × 1010,7.40 × 109,3.70 × 109,1.85 × 109 and 9.25 × 108 Bq/L,the cell inhibition ratios were (52.98±5.19)％,(46.29±2.80)％,(41.05±4.83)％,(33.68±3.79)％ and (17.89±2.78)％,respectively (F=33.882,P＜0.001).In the 4E5 group,when concentrations of 4E5 were 20.0,10.0,5.0,2.5 and 1.25 mg/L,the cell inhibition ratios were (32.98±3.99)％,(30.88±3.98)％,(27.14±2.05)％,(20.35±4.38)％ and (8.42±1.05)％,respectively.Accordingly,significantly higher growth inhibition rates for Raji cells than 4E5 at all antibody concentrations were tested (t=5.290,5.489,4.596,3.986 and 5.515,all P＜0.05).Conclusions The labeling efficiency and radiochemical purity of 131I-4E5 using Iodogen method is high,and the in vitro stability of 131I-4E5 is optimal.131I-4E5 can induce specific binding and cytotoxicity against Raji cells and may potentially serve as an agent for radioimmunoimaging and radioimmunotherapy of B cell lymphoma. Key words: Antibodies,monoclonal; Isotope labeling; Lymphoma,B-cell; Iodine radioisotopes; Radioimmunotherapy
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