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我们用Iodogen法标记McAb,体会到要想获得满意的标记效果,应控制好投料、pH、掌握反应时间及贮存时间几个环节。 材料和方法 一、材料: (一)Iodogen:化学名称为1,3,4,6—四氯,3α6α-二苯甘脲,分子量430,美国piere化学公司产品。用二氯甲烷(Sigma公司)溶解,分别配成50μl含3μg,5μg,10μg,25μg,50μg及100μg的浓度,取50μl涂布于2ml玻璃小瓶底部,用氮气轻轻吹干,当即盖上胶盖、封口,-20℃冻存备用。 (二)McAb BAC5:取液氮中冻存的杂交瘤细胞BAC5,复苏后培养至对数生长期,注射3.5×10~5个细胞/0.5ml至6～8周龄的BALB/c经产母鼠腹腔,10～14天后收集腹水。纯化腹水得纯度为93％的抗低分化鼻咽癌单抗BAC5,属IgG2a亚类,亲和力常数(Ka)值为8×10~9L·M~(-1),分子量149KD。 (二)无载体Na~(125)J,中国北京原子能中、研究院提。
We use the Iodogen method labeled McAb, experience to get a satisfactory labeling effect, should control the feed, pH, to master the reaction time and storage time of several links. Materials and methods First, the material: (a) Iodogen: chemical name 1,3,4,6-tetrachloro, 3α6α-diphenyl glycoluril, molecular weight 430, the United States piere chemical products. Dissolve with methylene chloride (Sigma) and make them into 50μl containing 3μg, 5μg, 10μg, 25μg, 50μg and 100μg respectively. Apply 50μl to the bottom of 2ml glass vial and gently blow dry with nitrogen. Cover, seal, -20 ℃ frozen spare. (B) McAb BAC5: BAC5 hybridoma cells cryopreserved in liquid nitrogen, cultured to logarithmic growth after the recovery, injected 3.5 × 10 ~ 5 cells / 0.5ml to 6 to 8 weeks of age BALB / c production Maternal abdominal cavity, 10 to 14 days after collecting ascites. Purified ascites was purified to 93% purity anti-poorly differentiated nasopharyngeal carcinoma monoclonal antibody BAC5, which belongs to IgG2a subclass. The affinity constant (Ka) value is 8 × 10-9L · M -1 and the molecular weight is 149KD. (B) without carrier Na ~ (125) J, Beijing, China Atomic Energy Research Institute mentioned.