抗人肺腺癌单克隆抗体—天花粉蛋白免疫毒素对荷人肺…

通过抗人肺腺癌单抗ＣＭＵ１５Ａ与天花粉蛋白的结合物即免疫毒素ＣＭＵ１５Ａ－ＴＣＳ对荷人肺腺癌裸鼠的导向治疗，观察了此免疫毒素在动物体内对实体瘤的导向性及靶向抑癌效果。结果：＾９９ｍＴｃ标记单抗经腹腔注射后２４ｈ在实体瘤内出现明显的靶向浓聚；此免疫毒素具有明显的靶向抑癌效果，腹腔用药和肿瘤内用药的抑瘤率分别为７６％和９９．４％；且可明显延长荷瘤裸鼠的生存时间；在整个疗程中未见明显的毒副作用发生。为免     

^99mTc标记卵巢癌单抗片段F（ab‘）2放射免疫显像的实验研究

目的：为了改进放射免疫显像（ＲＩＩ），进行＾９９ｍＴｃ标记卵巢癌单克隆抗体ＣＯＣ１８３Ｂ－Ｆ（ａｂ’）２ＲＨ的研究。方法：采用无茶果酶制单抗ＣＯＣ１８３Ｂ２片段（ａｂ’）２，改进ＳｎＣｌ２法进行＾９９ｍＴｃ标记片段Ｆ（ａｂ’）２及正常鼠ＩｇＧ（ＮＭＩｇＧ）；＾９９ｍＴｃ－ＣＯＣ１８３Ｂ２或＾９９ｍＴｃ－ＮＭＩｇＧ标记物分别经腹腔注入荷入卵巢癌裸鼠体内，１８ｈ进行ＲＩＩ。显像后测定并计算肿瘤与非肿瘤     

^99mTc—RBC的X射线光电子能谱研究

＾９９ｍＴｃ－标记的红细胞（＾９９ｍＴｃ－ＲＢＣ）是目前最常用的血池显像剂。本工作用体内标记法制备了＾９９ｍＴｃ－ＲＢＣ，采用Ｘ－射线光电子能谱（ＸＰＳ）对＾９９ｍＴｃ－ＲＢＣ中Ｔｃ原子的化学状态及其变化进行了测定。＾９９ｍＴｃ－ＲＢＣ于标记后１ｈ与标记后２ｈ所测ＸＰＳ能谱显示，表明＾９９ｍＴｃ－ＲＢＣ的稳定性佳。从ＲＢＣ与＾９９ｍＴｃ成键的情况看，多肽键上带负电荷的羧酸根氧原子与＾９９ｍＴｃ配合     

^99mTc—PYM的制备,动物实验及临床研究

研制适于临床肿瘤阳性显像的标记产品＾９９Ｔｃ－ＰＹＭ。方法：在原工作基础上，提高＾９９ｍＴｃ－ＰＹＭ的放射性强度，并通过注射前往产品中加入维生素Ｃ，提高肿瘤对＾９９ｍＴｃ－ＰＹＭ的摄取百分率。结果：定量判断肺部恶性病灶的灵敏度，特异性和准确性分别为９２％，１００％和９３％。＾９９ｍＴｃ－ＰＹＭ显像可用于肺癌诊断及分期，具有对疗效评价及预后判断的能力，该方法简便，安全，值得临床进一步推广应用。     

^99mTc标记抗CEA单抗放射免疫显像的临床研究

目的：采用放射性核素＾９９ｍＴｃ标记抗癌胚抗原（ＣＥＡ）抗体Ｃ５０进行放射免疫显像（ＲⅡ）以探讨其临床应用价值。方法：①对１５２例临床怀疑为各种肿瘤的患者做了ＲⅡ检查,包括卵巢肿瘤１１５例,肠道肿瘤２６例和肺肿瘤１１例。②用２－巯基乙醇还原法标记＾９９ｍＴｃ－Ｃ５０,其标记率达９０％以上,单抗用量１￣１．５ｍｇ。③静注地塞米松防过敏,＾９９ｍＴｃ－Ｃ５０静泳注射后４￣６ｈ采集平面或断层图像。④以病     

ＣＤ４５单抗介导的淋巴瘤荷瘤裸鼠三步法预定位放免显像

摘 要： 【目的】 评价生物素化ＣＤ４５单抗介导的９９ｍＴｃ－生物素在人Ｒａｊｉ细胞移植瘤裸鼠模型中三步法预定位放免显像的价值。【方法】 ＣＤ４５单抗及ＤＴＰＡ－生物素的９９ｍＴｃ标记采用直接标记法。取人Ｒａｊｉ细胞移植瘤裸鼠１８只,随机分为２组,每组９只。实验组为三步法预定位放免显像组,静注生物素化ＣＤ４５单抗１００ μｇ,４８ ｈ后静注亲和素２００ μｇ,再过４８ ｈ静注９９ｍＴｃ－生物素７．４ ＭＢｑ（２０ μｇ）;对照组为９９ｍＴｃ－ＣＤ４５单抗放免显像组,静注９９ｍＴｃ－ＣＤ４５单抗１００ μｇ。上述两组裸鼠分别于注药后３、６、１２ ｈ分别进行ＳＰＥＣＴ显像,每个时间点各取３只裸鼠断颈处死后,取脏器组织及肿瘤,称重后在γ计数仪中测量放射性计数,经放射性衰变校正后计算各脏器的％ＩＤ／ｇ及肿瘤／非肿瘤（Ｔ／ＮＴ）比值。【结果】平均每分子ＣＤ４５单抗约结合１２分子生物素,ＣＤ４５单抗及ＤＴＰＡ－生物素的９９ｍＴｃ标记率分别＞７０％和＞８０％。三步法给药后荷瘤裸鼠ＳＰＥＣＴ显像及生物分布示：整个显像期间血池内放射性均较低,肝脾见较多放射性浓聚;注射标记物后３－６ｈ,肿瘤显影清晰,并持续到１２ ｈ;注药后３、６和１２ ｈ肿瘤的％ＩＤ／ｇ分别为１．７３ ± ０．２２、１．２４ ± ０．０３和０．９４ ± ０．０７;肿瘤／血液比值分别为３．５、４．９和７．８;肿瘤／肌肉比值分别为８．２、８．９和１０．４。而静注９９ｍＴｃ－ＣＤ４５单抗后则可见肝脾及肾脏明显放射性聚集,１２ｈ血池内见较多放射性分布,肿瘤部位见有少量放射性集聚,１２ ｈ肿瘤的％ＩＤ／ｇ为０．８９ ± ０．１３,肿瘤／血液及肿瘤／肌肉的比值分别为１．６和２．５。【结论】与９９ｍＴｃ－ＣＤ４５单抗相比较,９９ｍＴｃ－生物素三步法预定位放免显像明显改善肿瘤Ｔ／ＮＴ比值,标记物注射后３ ｈ即可使肿瘤显影。     

抗人肺腺癌单克隆抗体—天花粉蛋白免疫毒素对荷人肺腺癌裸鼠模型的导向治疗

通过抗人肺腺癌单抗ＣＭＵ１５Ａ与天花粉蛋白的结合物即免疫毒素ＣＭＵ１５Ａ—ＴＣＳ对荷人肺腺癌裸鼠的导向治疗，观察了此免疫毒素在动物体内对实体瘤的导向性及靶向抑瘤效果，结果：￣（９９ｍ）Ｔｃ标记单抗经腹腔注射后２４ｈ在实体瘤内出现明显的靶向浓聚；此免疫毒素具有明显的靶向抑瘤效果，腹腔用药和肿瘤内用药的抑瘤率分别为７６％和９９．４％；且可明显延长荷瘤裸鼠的生存时间；在整个疗程中未见明显的毒副作用发生。为免疫毒素的临床应用提供了重要的基础资料。     

99mTc标记卵巢癌单抗片段F(ab'''')2 放射免疫显像的实验研究

目的：为了改进放射免疫显像（ＲＩＩ），进行９９ｍＴｃ标记卵巢癌单克隆抗体ＣＯＣ１８３Ｂ２Ｆ（ａｂ’）２ＲＩＩ的研究。方法：采用无花果酶制备单抗ＣＯＣ１８３Ｂ２片段Ｆ（ａｂ’）２，改进ＳｎＣｌ２法进行９９ｍＴｃ标记片段Ｆ（ａｂ’）２及正常鼠ＩｇＧ（ＮＭＩｇＧ）；９９ｍＴｃＣＯＣ１８３Ｂ２或９９ｍＴｃＮＭＩｇＧ标记物分别经腹腔注入荷人卵巢癌裸鼠体内，１８ｈ进行ＲＩ。显像后测定并计算肿瘤与非肿瘤组织放射性活度比值（Ｔ／ＮＴ）。结果：（１）无花果酶可制备ＣＯＣ１８３Ｂ２单抗片段Ｆ（ａｂ’）２；（２）改进ＳｎＣｌ２法进行９９ｍＴｃ标记Ｆ（ａｂ’）２，标记率达９０％以上；（３）１８ｈ后ＲＩ，可见９９ｍＴｃＣＯＣ１８３Ｂ２组荷瘤裸鼠较９９ｍＴｃＮＭＩｇＧ对照组移植瘤图像清晰，且Ｔ／ＮＴ值明显高。结论：应用片段Ｆ（ａｂ’）２可改善ＲＩＩ。     

^99mTc标记卵巢癌单抗的实验研究

用＾９９ｍＴｃ标记卵巢癌单抗，对标记方法，纯化手段，还原后抗体存时间与标记率关系，标记物免疫活性以及用该标记物对荷瘤裸鼠放免显象进行探讨，提示＾９９ｍＴｃ标记卵巢癌单抗的可行性。     

~(99m)Tc-MIBI显像和CA15-3联合检测对乳癌诊断的临床研究(摘要)

９９ｍＴｃ－ＭＩＢＩ显像和ＣＡ１５－３联合检测对乳癌诊断的临床研究（摘要）研究生汤学林导师张文莉周克敏吴光瑛（昆明医学院第三附属医院放疗科，昆明６５０１０６）关键词乳腺肿瘤，放射性核素显像，９９ｍＴｃ－ＭＩＢＩ，肿瘤相关抗原，ＣＡ１５－３中图分类号Ｒ...     

99mTc标记结肠癌单克隆抗体片段CEA．Fab的研制方法

目的　探讨一种高效便捷的99mTc CEA Fab标记化合物的研制方法。方法　采用 2 亚氨噻酚盐酸盐修饰CEA Fab ,通过GH转换 ,将99mTc标记在结肠癌单克隆抗体CEA Fab上。结果　已修饰抗体的标记率为 93.5 %,99mTc胶体含量为 2 .5 %,标记化合物有良好的稳定性和免疫活性。结论　成功地建立了结肠癌单抗片断CEA Fab的99mTc标记方法 ,该技术可以应用于其他抗体片段标记领域。     

99Tcm直接法标记Angiostatin及其在荷瘤小鼠体内研究

目的:99Tcm直接法标记血管抑素(angiostatin,AS),观察其在荷瘤小鼠体内的生物分布并进行显像,以探讨其在监测肿瘤对抗血管生成治疗的应答中的价值. 方法:氯化亚锡还原法进行AS的99Tcm标记,纸层析法测标记率;用人脐静脉内皮细胞系(ECV304)观察其生物活性;为了明确99Tcm-AS与肿瘤的结合,是否存在受体特异性,我们进行了封闭实验. 在给药前2 h用未标记AS预处理荷乳腺癌EMT6瘤株的BALB/c小鼠,尾静脉注射99Tcm-AS,观察其体内分布并进行显像. 结果:氯化亚锡还原法标记AS可获得较高的标记率(>95%),其抑制内皮细胞生长的生物活性与AS相似. 在荷瘤小鼠体内分布结果显示:肿瘤的摄取率随着时间延长而增加,在注射99Tcm-AS后2～12 h,肿瘤可清晰显像,同时,用未标记AS预先封闭肿瘤可使肿瘤对99Tcm-AS摄取率下降. 结论:99Tcm-AS在荷瘤小鼠体内可浓聚于肿瘤,肿瘤对显像剂的摄取存在一定的特异性;99Tcm-AS有可能在肿瘤抗血管生成治疗疗效评价中发挥作用.     

99Tcm-annexinV在肺癌化疗评估中的应用

目的制备放射性药物99Tcm-annexinV,评估其在早期预测肺癌化疗效果中的作用。方法通过毕赤酵母重组表达、硫酸铵沉淀和分子筛层析纯化得到annexinV,采用氯化亚锡还原法在annexinV的氨基端标记放射性核素99Tcm,经脱盐柱纯化获得标记产物。采用薄层层析测定99Tcm-annexinV的标记率和放射性化学纯度,外露磷脂酰丝氨酸的红细胞测定99Tcm-annexinV的生物活性。通过在615小鼠腋部皮下接种LA795细胞和组织插块法获得荷肺癌小鼠模型,经腹腔注射环磷酰胺治疗肺癌后6、12、24和48h测定99Tcm-annexinV在小鼠体内的分布情况。结果毕赤酵母工程菌株分泌表达annexinV,经硫酸铵分级沉淀和分子筛层析纯化后annexinV得以有效回收。室温下30min完成anne-xinV的99Tcm标记,标记率为50.2%。99Tcm-annexinV放射性化学纯度为93.9%,其生物活性保存良好。生物分布实验表明,99Tcm-annexinV通过肾脏排泄。615荷肺癌小鼠模型经过环磷酰胺化疗后48h,99Tcm-annexinV在肿瘤组织的摄取达到最高,肿瘤/肌肉放射性摄取率之比为6.34,肿瘤/血液放射性摄取率之比为4.09。结论制备了毕赤酵母来源的99Tcm-annexinV,有可能应用于早期预测肺癌化疗效果。     

~(99)Tc~m-EC-MN鼻咽癌乏氧显像的实验研究

目的 通过对99Tcm-EC-MN在鼻咽癌体外及体内模型中生物学分布的研究,探讨99Tcm-EC-MN肿瘤乏氧显像的临床意义.方法 利用99TcmO4标记EC-MN,纸层析法测定标记率.99Tcm-EC-MN分别加入正常条件和乏氧条件下培养的鼻咽癌细胞株内,在不同时间段检测99Tcm-EC-MN在细胞内的分布差别.建立鼻咽癌裸鼠肿瘤模型,自荷瘤鼠尾静脉注入99Tcm-EC-MN 3.7 MBq(0.1 ml)后0.5、1、2、4、6和8 h分别进行平面显像,观察99Tcm-EC-MN的体内分布情况.在各时相分别处死荷瘤裸鼠,取各脏器组织标本、称重并测量放射性计数,计算各标本每克组织百分注射剂量率(%ID/g).对分离出的瘤体,应用CD34单克隆抗体及血管内皮生长因子(VEGF)单克隆抗体进行免疫组化染色.结果 正常条件和乏氧条件下培养的鼻咽癌细胞株对99Tcm-EC-MN的吸收差异有高度统计学意义(P<0.01).所有荷瘤裸鼠的肿瘤均呈阳性显像,静脉注射后0.5 h肿瘤灶即显影,4 h时显影最清晰.肿瘤/肌肉比值1、2、4 h分别达2.24倍、4.75倍、6.41倍.99Tcm-EC-MN在正常组织中清除快,主要经泌尿系统及肠道排出.免疫组化结果和放射性计数结果具有显著相关性(r>0.9,P<0.01).结论 99Tcm-EC-MN作为一种乏氧显像剂,具有良好的开发前景.     

Diagnostische Bedeutung von pentavalenter 99m Tc-Dimercaptobern-steinsaure ([V]-DMSA) bei Kopf-Hals-Tumoren

The purpose of this investigation was to assess the biodistribution of 99m Tc-[V]-DMSA in human being,and its diagnostic value in patients with head and neck tumours.20 patients with histologically confirmed head and neck tumours were examined with planar as well as SPECT scintigraphy. Whole body scintigraphy in different time after injection of 99m Tc-[V]-DM-SA was performed to assess the major sequential organ biodistribution. Our results showed that the blood clearance of 99m Tc-[V]-DMSA was bi-exponential. All organs except kidneys showed a relatively rapid elimination of 99m Tc-[V]-DMSA. The kidneys showed a increasing accumulation in the first 2h, which is probably due to the tubular mabsorption of 99m Tc-[V]-DMSA. In 15 of 20 patients, 19 lesions could be proven by means of planar scintigraphy (corresponding sensitivity of 75%). 29 lesions in 18 patients could however be detected by the application of SPECT (corresponding sensitivity of 90%). Except primary tumours and local lymphadenmetastases in 5 patients distant metastases (3 thorax wall,I liver and I ingvinal/paravesicle) were found. Altogether a sensitivity of 76.9% and a specificity of 71.4% were calculated for the detection of primary tumour. The sensitivity and specificity for the exploration of lymphadenmetastases were 75% and 100%. In conclusion, this study shows that 99m Tc-[V]-DMSA, particularly with SPECT imaging, is useful in localising the primary tumours and lymphadenmetastases as well as distant metastases of head and neck tumours. The possible therapeutic application of 188/186Re-[V]-DMSA is also evaluated.     

Abdominal scanning using autologous human red blood cells labeled with Tc-99m: detection of a horseshoe kidney

A horseshoe kidney was incidentally diagnosed while performing an abdominal scan using autologous human red blood cells labeled with Tc-99m (Tc-99m RBC) on a patient with suspected gastrointestinal bleeding. This patient repeatedly tested positive on occult blood stool examinations. In the first scintigram, an increase in well-defined activity was found on the right side under the liver. This could have been caused either by bleeding of the gastric antrum or the duodenal bulb. A second abdominal scan with Tc-99m RBC revealed faint radionuclide accumulation in the shape of a horseshoe with symmetrical, well-defined activity. However, the well-defined activity decreased with time on both images, and on this basis a horseshoe kidney was diagnosed.     

99mTc标记葡萄糖99mTc-EC-DG在正常小鼠体内的分布研究

目的研究^99mTc标记葡萄糖99mTc-EC-DG在正常小鼠体内的生物学分布规律,用于临床肿瘤显像作基础研究.方法正常昆明小鼠48只,分8组,每组6只,实验前小鼠禁食8 h以上.尾静脉注射^99mTc-EC-DG 3.7 MBq (100 μCi) 后5 min、15 min、30 min、1 h、2 h、4 h、8 h和24 h放血处死,取心、肝、脾、肺、脑、肾、肌肉、骨、小肠、胃和血液等组织或器官,称重并测量其放射性,计算每克组织百分注射剂量率(%ID/g).结果^ 99mTc-EC-DG主要经肾脏代谢,脑不吸收^99mTc-EC-DG,肌肉组织摄取亦较少.各组织、器官的百分剂量率在1 h内除血液下降稍慢外,其余均有明显下降.结论^ 99mTc-EC-DG标记方便,体内外稳定性好,小鼠体内生物学分布表明其可能是一种较好的葡萄糖代谢显像剂.     

NHS-MAG3为螯合剂的99mTc-寡核苷酸标记特性

目的 探讨以NHS-MAG,作为螯合剂的^99mTc-寡核苷酸(^99mTc-ON)的标记特性。方法将NHS-MAG,与长度为15个碱基的c-myc mRNA的反义(ASON)、正义(SON)和无义寡核苷酸(MON)偶联,进行^99mTc标记,Sephadex G25 柱层析分离纯化,评价^99mTc-ON的标记率、放化纯和稳定性;将ON-MAG,偶联物置-20℃贮存15 d、1月、2月后进行^99mTc标记,观察标记率的变化,评价ON-MAG3的稳定性;用三氯乙酸沉淀法测定^99mTc-ON的血浆蛋白结合率。结果 ^99mTc-ASON、^99mTc-SON和^99mTc-MON的标记率分别为68．41％、66．24％和69．38％,纯化后放化纯分别为96．98％、95．34％和94．62％。三者在室温和血清中均较稳定。ON-MAG3在-20℃保存2月后,标记率未见明显下降。三种^99mTc-ON的血浆蛋白结合率均小于13％。结论以NHS-MAG3作为螯合剂的^99mTc-ON具备优良的标记特性,标记率、放化纯较高,稳定性好,血浆蛋白结合率低,是一种有临床应用潜力的放射性标记药物。     

卵巢癌荷瘤裸鼠~(99m)TcCOC183B_2放射免疫显像结果分析

目的 :研究99mTc标记抗卵巢上皮癌单克隆抗体COC183B2 在荷瘤裸鼠体内的分布 ,并进行放射免疫显像 ,为临床应用提供依据。方法 :COC183B2 用预亚锡法标记后 ,经腹腔注入荷瘤裸鼠体内 ,在 2 4小时进行放射免疫显像 ,并观察裸鼠体内的生物学分布。结果 :与对照组相比 ,标记抗体在肿瘤组织中出现了特异性浓聚。获得清晰的显像 ,T/NT比值在 0 99～ 5 99之间 ,T/B比值为 1 19。结论 :说明COC183B2 可特异地与卵巢癌结合 ,具有导向卵巢癌的作用     

Pre-clinical evaluation of a new indirectly labeled 99mTc-6-hydrazinopyridine-3-carboxylic acid (HYNIC)-depreotide with HYNIC as bifunctional chelator

Background  Technetium-99m or ^99mTc is widely used for labeling peptide in nuclear medicine. Somatostatin and its analog can inhibit tumor cell growth after binding with its receptor. This research was to study the preclinical effect of a new ^99mTc-6-hydrazinopyridine-3-carboxylic acid (HYNIC)-depreotide, indirect ^99mTc labeling of depreotide using HYNIC as a bifunctional chelator.

Methods  The cyclopeptide, cyclo-[(N-Me) Phe-Tyr-D-Trp-Lys-Val-Hcy], the linear peptide, and [ClCH₂-CO×b-Dap-Lys- Cys-Lys×amide] were synthesized by Fmoc solid-phase synthesis. The cyclopeptide and the linear peptide were linked by liquid-phase synthesis. The product depreotide was isolated and purified by high performance liquid chromatography and was confirmed by mass spectrography. Depreotide was labeled with ^99mTc through a direct labeling method, using HYNIC as a bifunctional chelator. Paper chromatography method was used to calculate the labeling rate, and through the comparative analysis selected the best mark conditions. The new ^99mTc-HYNIC-depreotide was tested by high-performance liquid chromatography (HPLC). The internalization and externalization rates of the new ^99mTc-HYNIC-depreotide were studied in A549 cells. Furthermore, biodistribution of the radiopeptide was studied in nude mice, bearing tumors from human lung carcinoma cells SPC-A1.

Results  The molecular of synthesize depreotide was 1358, and the purity of it was 95.29%. The labeling efficiency of ^99mTc-HYNIC-depreotide was highest at pH 6.0 and 15°C, about (70.95±0.84)%. The labeling rate of the new ^99mTc-HYNIC-depreotide rose to a peak of (20.75±0.48)% at 60 minutes in A549 cells at 37°C and decreased slightly later, while it elevated gradually during the time course at 4°C and 25°C. The internalization rate of the new ^99mTc-HYNIC-depreotide at 37°C increased gradually and reached the peak of 84.4% in 120 minutes, while the externalization rate of the new ^99mTc-HYNIC-depreotide was always less than 20%. In mice bearing the experimental SPC-A1 tumor, the new ^99mTc-HYNIC-depreotide demonstrated a high tumor uptake of (4.05±0.04)% ID/g at 1.5 hpi and remained high ((2.51±0.06)% ID/g) at 4 hpi. The tumor-to-lung activity concentration ratio (T/Lu) was very high for the new ^99mTc-HYNIC-depreotide at all time points. So did the tumor-to-muscle activity (T/Mu) and tumor-to-blood activity concentration ratios (T/Bl).

Conclusion  The findings suggested that the new ^99mTc-HYNIC-depreotide might be a promising candidate radiopharmaceutical for imaging somatostatin receptor positive lung cancer.