Targeting therapy against neuroendocrine tumor employing anticalcitonin gene antibody

1.裸鼠移植我们用甲状腺髓样癌组织作神经内分泌肿瘤。虽然我们最近进行了三例甲状腺髓样癌的移植，但最终都失败了。结论：1.甲状腺髓样癌的移植非常困难。分离的兔Ig组分血清与DEAE Sephadex A-50混合，用0.1M Tris HCl(ph8.0)预校准，在4℃下搅拌1小时，过滤后，用5ml缓冲液洗涤凝胶。重复吸附过程，沉淀出组合滤液。兔抗人免疫球蛋白Fab和Fc片段的分离用木瓜蛋白酶切割，用Protein A-SepharoseCL-4B蛋白A-SepharoseCL-4B柱分离兔抗人Ig G，在含有半胱氨酸和NaEDTA的1ml PBS中加入等量的木瓜酶，37℃孵育3-5小时，加入碘乙酸后，透析液与PBS过夜。兔抗人PDN-21免疫电泳组分在琼脂糖凝胶中溶解于乳酸钙缓冲液中，140V作用45min。以氯胺T为氧化剂，用~(125)I标记人PDN-21。5.血清PDN-21、CGRP、降钙素的测定采用我们的方法测定血清PDN-21、CGRP、降钙素的水平。

1. Transplantation gainst nude mouse We used tissue of medullary thyroid carcinoma sa a neuroendocrine turmor. Although we transplanted three cases of medullary thyroid carcinoma recently, all of them failed for transplantation finally. Transplantaion of medullary thyroid carcinoma is very difficult.2. Isolation of Rabbit Ig Fraction Serum was mixed with DEAE Sephadex A-50, preequiblibrated with 0.1 M Tris Hcl(ph8.0) and stirred for 1 hr at 4 C. After filtration the gel was washed with 5ml of the buffer. The adsorption procedure was repeated, and the combined filtrated ware precipitated.3. Separation of Fab and Fc Fragment Pufified rabbit antihuman IgG was cleaved by papain and the Tab and Fc fragments separated using a protein A-Sepharose CL-4B colum Rabbit IgG in 1 ml PBS containing cystein and NaEDTA received papain of the amount of IgG and was incubated for 3-5 hr at 37 C. After addition of iodoacetic acid, the soulution was dialysed overnight against PBS.4. Immunoelectophoresis of Rabbit Antihuman PDN-21 IgG Fraction Electrophoresis was carried out in agarose gel dissolved in calcium lactate buffer for 45 min using 140 volts. Human PDN-21 was labeled with 125I using chloramine-T as oxidizing agent. Electrolytic iodination of anti-PDN-21 IgG fraction was carried out in PBS using radioactive iodine.5.Measurement of serum PDN-21, CGRP, calcitonin Serum levels of PDN-21, CGRP and calcitonin were measured by the method by us.