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Luo XF, Zhang B, Ma HB, Tang L, Zhou X, Xu Y, Wu YW, Zhang XG: [Preparation of iodine-125 labeled anti-human CD40 monoclonal antibody 5H6 and its binding properties in ovary cancer cell line HO8910 in vitro]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Jun;24(6):590-3.
AIM: To prepare iodine-125 labeled anti-human CD40 monoclonal antibody 5H6 ((125) I-5H6) and investigate the binding properties of (125) I-5H6 to HO8910 cells in vitro. METHODS: The mAb 5H6 was labeled with Na (125) I using chloramine-T method. The labeling efficiency and radiochemical purity of (125) I-5H6 were measured by paper chromatography. The stability of (125) I-5H6 in vitro was monitored by trichloroacetic acid (TCA) precipitation. The dissociation constant (K (d)) of (125) I-5H6 from HO8910 cells and the numbers of maximum binding sites (B (max)) were obtained by Scatchard analysis. Immunoreactivity of (125) I-5H6 to HO8910 cells was determined by "Lindmo" assay and its improved method. Internalization and retention of (125) I- 5H6 by HO8910 cells were studied by cell binding experiments. RESULTS: (1) The labeling efficiency of (125) I-5H6 was (85.4+/-5.2)% and its radiochemical purity was (99.2+/-0.5)%. (2) Radiochemical purity of (125) I-5H6 was (80.3+/-4.7)% after 7 days in 4 Degrees Celsius PB (phosphate buffer) and (95.3+/-0.8)% after 24 hours in 37 Degrees Ceslius plasma. (3) At 4 Degrees Celsius, K (d) for (125) I-5H6 to HO8910 cells was (0.711+/-0.06) nmol/L and Bmax was (2.17+/-0.08) x10 (5) sites/cell. (4) Immunoreactivity of (125) I-5H6 was (38.6+/-5.4)%. (5) Retention rate of (125) I-5H6 with HO8910 cells was (89.8+/-6.0)% after 2 hours at 4 Degrees Celsius and internalization rate of (125) I-5H6 by HO8910 cells was (54.9+/-2.6)% after 2 hours at 37 Degrees Celsius. CONCLUSION: (125) I-5H6 possesses good labeling efficiency and perfect radiochemical purity. (125) I-5H6 has suitable stability in vitro and its immunoreactivity was not low. The high affinity of (125) I-5H6 with HO8910 cells will benefit further animal study in vivo. |
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