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抗人卵巢癌COC183B2/抗CD3单链双特异抗体的构建和表达 被引量:2
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作者 赖娟 李小平 +3 位作者 冯捷 程洪艳 姚煜 叶雪 《北京大学学报(医学版)》 CAS CSCD 北大核心 2002年第4期358-361,共4页
目的 :构建和表达一种可与卵巢癌细胞和淋巴细胞结合的双特异抗体。方法 :利用PCR分别扩增抗人CD3单链抗体 (singlechainvariablefragment,ScFv)重链可变区 (variableregionoftheheavychain ,VH)和轻链可变区 (variableregionofthelight... 目的 :构建和表达一种可与卵巢癌细胞和淋巴细胞结合的双特异抗体。方法 :利用PCR分别扩增抗人CD3单链抗体 (singlechainvariablefragment,ScFv)重链可变区 (variableregionoftheheavychain ,VH)和轻链可变区 (variableregionofthelightchain ,VL) ,重组抗人CD3ScFv ,经测序后将其克隆入有链间连接肽基因序列的载体pALM中 ,再将抗人卵巢癌COC183B2ScFv克隆至紧邻链间连接肽前形成COC183B2 /抗CD3单链双特异抗体(single-chainbispecificantibody ,scBsAb)的重组。最后将COC183B2 /抗CD3scBsAb克隆入表达载体 pTMFC中进行表达 ,同时分别表达抗人卵巢癌单抗COC183B2和抗人CD3单抗的ScFv作为对照组。用ELISA、流式细胞学方法和玫瑰花环实验对scBsAb进行免疫学活性测定。结果 :成功构建抗人卵巢癌COC183B2 /抗CD3scBsAb ;其表达的蛋白链相对分子质量约 6 0 ;ELISA结果显示scBsAb可与抗原OC183B2结合 ,流式细胞学结果显示scBsAb可与抗原CD3结合 ,花环实验显示scBsAb在体外可引导效应细胞聚集在靶细胞周围。结论 :构建和表达抗人卵巢癌COC183B2 /抗CD3scBsAb成功 ,且具有与抗原OC183B2。 展开更多
关键词 卵巢癌 coc183b2 双特异抗体 淋巴细胞 抗CD3单链抗体 PCR
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Evaluation of COC183B2 antibody targeting ovarian cancer by near-infrared fluorescence imaging 被引量:4
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作者 Chen Zhang Xinyu Ling +10 位作者 Yanxiu Guo Cunzhong Yuan Hongyan Cheng Xue Ye Ruiqiong Ma Yinli Zhang Yi Li Xiaohong Chang Beihua Kong Tao Liu Heng Cui 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 2019年第4期673-685,共13页
Objective: To evaluate the imaging potential of a novel near-infrared(NIR) probe conjugated to COC183 B2 monoclonal antibodies(MAb) in ovarian cancer(OC).Methods: The expression of OC183 B2 antigen in OC was determine... Objective: To evaluate the imaging potential of a novel near-infrared(NIR) probe conjugated to COC183 B2 monoclonal antibodies(MAb) in ovarian cancer(OC).Methods: The expression of OC183 B2 antigen in OC was determined by immunohistochemical(IHC) staining using tissue microarrays with the H-score system and immunofluorescence(IF) staining of tumor cell lines.Imaging probes with the NIR fluorescent dye cyanine 7(Cy7) conjugated to COC183 B2 Mab were chemically engineered. OC183 B2-positive human OC cells(SKOV3-Luc) were injected subcutaneously into BALB/c nude mice. Bioluminescent imaging(BLI) was performed to detect tumor location and growth. COC183 B2-Cy7 at 1.1,3.3, 10, or 30 μg were used for in vivo fluorescence imaging, and phosphate-buffered saline(PBS), free Cy7 dye and mouse isotype immunoglobulin G(IgG)-Cy7(delivered at the same doses as COC183 B2-Cy7) were used as controls.Results: The expression of OC183 B2 with a high H-score was more prevalent in OC tissue than fallopian tube(FT) tissue. Among 417 OC patients, the expression of OC183 B2 was significantly correlated with the histological subtype, histological grade, residual tumor size, relapse state and survival status. IF staining demonstrated that COC183 B2 specifically expressed in SKOV3 cells but not HeLa cells. In vivo NIR fluorescence imaging indicated that COC183 B2-Cy7 was mainly distributed in the xenograft and liver with optimal tumor-to-background(T/B)ratios in the xenograft at 30 μg dose. The highest fluorescent signals in the tumor were observed at 96 h postinjection(hpi). Ex vivo fluorescence imaging revealed the fluorescent signals mainly from the tumor and liver. IHC analysis confirmed that xenografts were OC183 B2 positive.Conclusions: COC183 B2 is a good candidate for NIR fluorescence imaging and imaging-guided surgery in OC. 展开更多
关键词 coc183b2 ANTIBODY NEAR-INFRARED FLUORESCENCE imaging OVARIAN CANCER
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^(99)Tc^m-COC183B_2用于卵巢癌荷瘤裸鼠放射免疫显像的实验研究
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作者 曲军英 闫平 +1 位作者 林好学 朱凤川 《中华核医学杂志》 CAS CSCD 北大核心 1999年第3期180-181,共2页
用放射性核素标记抗肿瘤抗体进行体内肿瘤放射免疫显像(RI),有较高的特异性和敏感性。目前国外允许用于卵巢癌诊断的单抗仅几种,而RI药盒仅Immun4(抗CEA),但其来源少,价格高,限制了临床使用。本研究用99T... 用放射性核素标记抗肿瘤抗体进行体内肿瘤放射免疫显像(RI),有较高的特异性和敏感性。目前国外允许用于卵巢癌诊断的单抗仅几种,而RI药盒仅Immun4(抗CEA),但其来源少,价格高,限制了临床使用。本研究用99Tcm标记国内制备的抗人卵巢上皮癌单... 展开更多
关键词 卵巢癌 放射免疫显像 coc183b2 实验研究
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双特异抗体COC183B_2×anti-CD28对卵巢上皮癌细胞杀伤效力的研究 被引量:1
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作者 顾莹 冯捷 +5 位作者 周志勇 张众 李小平 林晴 叶雪 黄华粱 《现代妇产科进展》 CSCD 2002年第4期241-244,I001,共5页
目的 :检测双特异抗体 (bispecificantibody ,BsAb)COC183B2 ×anti CD2 8在体外介导预激活的外周血单个核细胞 (PBMC)对卵巢上皮癌细胞系SKOV3的杀伤效力。方法 :在大肠杆菌中对用基因工程方法构建的COC183B2 ×anti CD2 8进... 目的 :检测双特异抗体 (bispecificantibody ,BsAb)COC183B2 ×anti CD2 8在体外介导预激活的外周血单个核细胞 (PBMC)对卵巢上皮癌细胞系SKOV3的杀伤效力。方法 :在大肠杆菌中对用基因工程方法构建的COC183B2 ×anti CD2 8进行诱导表达 ,用ELISA方法测定表达产物与卵巢癌抗原和CD2 8抗原结合的活性后 ,与SKOV3和预激活的PBMC共孵育。 6h后用非放射性细胞毒分析试剂测定预激活的PBMC对SKOV3细胞毒性作用 ,并在镜下观察SKOV3的形态学改变。结果 :ELISA结果显示 ,COC183B2 ×anti CD2 8可与卵巢癌抗原和CD2 8抗原双向结合 ;细胞毒性试验显示 ,在体外COC183B2 ×anti CD2 8能介导预激活的PBMC特异杀伤SKOV3细胞 ;4h时镜下可见玫瑰花环形成 ,12h可见肿瘤细胞溶解。结论 :COC183B2 ×anti CD2 展开更多
关键词 coc183b2×anti-CD28 卵巢上皮癌 双特异抗体 卵巢肿瘤 外周血单个核细胞 细胞毒性试验 生物治疗
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