单克隆抗体连接表阿霉素导向治疗胃癌的实验研究

作者首次报道了胃癌单克隆抗体MGb_2-表阿霉素结合物的制备及其对胃癌细胞的体外杀伤作用。首先用高碘酸钠氧化葡聚糖,使其成为多醛基葡聚糖,并以此为中间载体将表阿霉素与MGb_2连接。经测定,每一抗体能携带45个药物分子,且其抗体活性保存良好。该结合物对胃癌细胞SGC-7901杀伤作用优于游离药物及无关抗体结合物,且对非靶细胞(Hep-2)毒性作用很弱。说明表阿霉素-葡聚糖-MGb_2对胃癌细胞具有较强选择性杀伤作用,并可能成为胃癌导向治疗的免疫抗癌药物。     

胃癌单克隆抗体-丝裂霉素C结合物的制备及其细胞毒特征

作者报道胃癌单克隆抗体MG11-丝裂霉素C(MMC)结合物的制备及其对肿瘤细胞的杀伤作用。首先用戊二酐处理MMC,于MMC上引入羧基,MMC衍生物与N羟基琥珀酰亚胺及2,2′-二环已基碳二亚胺反应,得到MMC活性酯,后者与抗体反应将MMC引入抗体中。经测定,每克分子抗体中引入约6～7克分子药物,结合物对人胃癌细胞KATⅢ具有较强的选择性杀伤作用,在0.56μg/ml水平(药物浓度)对肿瘤细胞杀伤牢达60％,优于游离药物(51％)及无关抗体结合物(9.3％),提示选用的胃癌单抗对MMC具有较好的导向作用。     

抗人B淋巴细胞白血病导向药物生物活性的研究

抗肿瘤新药表阿霉素（ＥＰＩ）通过葡聚糖Ｔ１０与抗人Ｂ淋巴细胞白血病单抗（Ｂ１５９）连接制备免疫结合物。该结合物保持了原有的抗体活性，在体外对人Ｂ淋巴母细胞Ｒａｊｉ显示出特异的杀伤作用，并具有良好的稳定性。     

柔红霉素连接单克隆抗体用于胃癌导向治疗的实验研究

本文采用直接和间接偶连方法将柔红霉素成功连接于胃癌单克隆抗体MGb_2,每一抗体所载的药物分子分别为6个(直接法)和54个(间接法)。ELISA证实所制备结合物中MGb_2活性保存较好,且能与胃癌靶细胞特异地结合。噻唑蓝(MTT)比色法体外细胞毒试验表明结合物对胃癌细胞SGC—7901具有选择性杀伤作用,而对非靶细胞毒性很弱,提示柔红霉素对胃癌细胞杀伤作用是由单抗MGb_2介导的。     

胃癌单克隆抗体用于氯甲喋呤导向治疗的实验研究

作者报道胃癌单克隆抗体 MGb_2-BSA-MTX结合物的制备及其对肿瘤靶细胞的杀伤作用。首先将 MTX引入BSA,继与单抗连接得到结合物。结合物中IgG:BSA:MTX克分子比为1:15:62。偶联过程中抗体活性无明显丧失,结合物与肿瘤靶细胞的结合能力与未修饰抗体相近。噻唑蓝活细胞染色的检测结果表明,结     

人转铁蛋白-阿霉素结合物对耐阿霉素K562细胞的体外杀伤作用

目的 探讨阿霉素（ＡＤＭ）与人转铁蛋白（Ｔｆ）结合物对耐阿霉素Ｋ５６２细胞株的体外杀伤作用。方法 采用葡聚糖（ＤＥＸ）Ｔ－１０作为中介体、联接Ｔｆ与ＡＤＭ制备结合物Ｔｆ－ＡＤＭ,用ＭＴＴ法比较结合物对耐ＡＤＭ及ＡＤＭ敏感的Ｋ５６２细胞的体外杀伤作用强度。结果 结合物对耐ＡＤＭ的Ｋ５６２细胞株７２ｈ培养、ＭＴＴ法测定结合物对耐ＡＤＭ的Ｋ５６２细胞ＩＣ５０为４．９μｍｏｌ,游离ＡＤＭ的ＩＣ５０〉１７．２４μｍｏｌ,Ｔｆ对这两种细胞都没有体外杀伤作用。结论 结合物Ｔｆ－ＡＤＭ可提高ＡＤＭ对耐ＡＤＭ的Ｋ５６２细胞株的杀伤作用,此作用与Ｔｆ无明显关系。     

半乳糖结合位点封闭免疫毒素的研制及其杀伤活性

作者报道B链半乳糖结合位点封闭的蓖麻毒素-胃癌单克隆抗体MGb_2结合物的制备及其细胞毒特征。首先将引入碘乙酰基团的毒素与引入巯基的抗体反应,得到的结合物通过琼脂糖亲和层析柱除去仍保留半乳糖结合能力的部分。细胞毒性试验结果表明,结合物对肿瘤靶细胞具有较强的选择性杀伤作用,在1.0×10~(-11)mol/L水平,对人胃癌细胞KATOⅢ的杀伤率为46％,优于MGb_2-蓖麻毒素A链结台物(12％),而对非靶细胞作用很弱,在1.0×10~(-9)mol/L水平对正常人胚肺细胞SL_7杀伤率仅为42％,无需半乳糖拮抗。     

双功能抗体抗CD3/抗CyclinD1的制备及对子宫内膜癌细胞的杀伤作用

目的 :制备抗CD3/抗CyclinD1双功能抗体 ,探讨子宫内膜癌主动免疫治疗的可行性 .方法 :以化学交联方法将抗CD3单克隆抗体与抗CyclinD1单克隆抗体交联为双功能抗体 ,进行介导对子宫内膜癌细胞的杀伤作用 .结果 :该双功能抗体介导效应细胞对靶细胞的杀伤率为 6 1.2 3% ,高于单抗CD3(4 2 .15 % )的介导作用 (P <0 .0 1) ,并随着效靶比的提高杀伤率也升高 (78.96 % ) .结论 :采用化学交联法制备的抗CD3/抗CyclinD1双功能抗体具有主动免疫治疗宫颈癌的临床可行性 .     

CEA单抗-MTX结合物的选择性细胞毒作用

氨甲喋呤(MTX)和癌胚抗原(CEA)单克隆抗体(McAb)共价结合,制备成MTX-McAb结合物。反应中MTX/McAb为1∶40时,结合物中的MTX/McAb为1∶14,抗体蛋白无明显变性,结合物保留了相当的抗体特异性和药物细胞毒活性。细胞毒试验表明,MTX-McAb对CEA阳性大肠癌细胞HT-29的细胞毒作用明显大于NIgG-MTX和MTX,且能被CEA McAb竞争性阻断。对CEA阴性细胞,MTX-McAb,NIgG-MTX和MTX三者的细胞毒作用无明显差异。提示MTX-McAb通过CEAMcAb的导向作用,能选择性地杀伤CEA阳性细胞。CEA McAb可作为MTX等化疗药物的载体用于CEA阳性肿瘤的治疗。     

高效价抗人血红蛋白单克隆抗体的制备及鉴定

目的制备高效价抗人血红蛋白(Hb)单克隆抗体并对其进行鉴定。方法用纯化人血红蛋白免疫Balb/c小鼠,取免疫后小鼠脾细胞与骨髓瘤细胞融合,筛选分泌抗人Hb单克隆抗体的细胞株,将阳性细胞株接种于小鼠腹腔制备单克隆抗体并对腹水抗体进行纯化,测定抗体亚类,分析其敏感性、特异性及纯化抗体的相对亲和力。结果共获得3株单克隆抗体杂交瘤细胞株,均与人Hb有较高的亲和力,其敏感性可达0.5μg/ml,而且与人肌红蛋白(Mb)及各种动物血红蛋白、肌红蛋白均无交叉反应。结论3株单克隆抗体杂交瘤细胞株有较好的特异性,与人Hb有较高的亲和力。     

抗人食管癌单克隆抗体和争光霉素A_6结合物的制备及其对食管癌细胞的细胞毒作用

研究目的探索食管癌导向化疗的途径研究设计用异双功能基试剂SPDP联接抗人食管癌单抗和争光霉素A_6,然后用MTT法测定该结合物对食管癌细胞的特异性细胞毒作用。研究单位新乡医学院基础医学部处理方法采用MTT法,通过测定细胞代谢活性来反映单抗—A_6结合物对食管癌细胞的特异性杀伤作用。靶细胞为食管癌细胞Eca8714,非靶细胞为宫颈癌细胞HL,设有结合物组和对照组(游离A_6)。结果结合物对食管癌细胞的杀伤率比游离争光霉素强10倍。0.1μM时即显示杀伤癌细胞活性。首次发现争光霉素A_6对宫颈癌细胞HL的杀伤率明显高于对食管癌细胞Eca8714的杀伤率。结论单抗和争光霉素A_6的结合物显示出较好的特异性细胞毒作用.为食管癌导向化疗迈出了有益的探索性的一步。     

单克隆抗体-苯并卟啉衍生物偶联物的制备及其在体外抗人肺癌的效应

作者用修饰-聚乙烯醇(M-PVA)作载体,通过1-乙基-3-(3-二甲基氨丙基)-碳化二亚胺盐酸盐(EDCI)将抗人肺癌单克隆抗体5E8和新颖的光敏药物苯并卟啉衍生物-一元酸-A环(BPD-MA)桥连成一偶联物。结果表明5E8偶联物除保留5E8抗体活性和BPD-MA杀细胞效应外,在体外还对人肺癌细胞有特异性杀伤作用。     

Targeting therapy with adriamycin-monoclonal antibody (HAb18) conjugate in nude mice with human hepatocellular carcinoma xenografts]

Adriamycin (ADM) was linked covalently to the murine monoclonal antibody (McAb) HAb18 against human hepatocellular carcinoma via a dextran T-40 bridge. The molar ratio of HAb18: DEX: ADM was 1:2.4:56 in the conjugate. It was confirmed by immunochemical staining and ELISA that the antigen binding capacity of HAb18 in the conjugate was well preserved. In vitro study indicated that the conjugate cytotoxicity to tumor target cell lines was selective. The radio-immunoimaging and distribution of the conjugate labelled with 125I in the nude mice bearing human hepatocellular carcinoma showed that 125I-HAb 18-DEX-ADM conjugate was localized in the tumor site. The HAb18-DEX-ADM targeting therapy in nude mice bearing human hepatocellular carcinoma showed the following results: decreased tumor size and partly necrosed cancer tissue in contrast to the rapidly growing tumor in the control group (P < 0.05). Our study suggested that targeting therapy with HAb18-DEX-ADM conjugate may be useful in the treatment of patients with hepatocellular carcinoma.     

消痰散结方对MKN-45人胃癌细胞黏附分子CD_(44)V_6表达的影响

目的 :观察消痰散结方对体外培养的 MKN- 45人胃癌细胞黏附分子 CD44 V6表达的影响 ,初步探讨了消痰散结方抑制胃癌细胞转移的作用环节。方法 :制备消痰散结药物血清 ,体外培养 MKN- 45人胃癌细胞 ,拟消痰散结方药物血清干预培养细胞 ,采用免疫组化 ABC法检测胃癌细胞中 CD44 V6的表达。结果 :中药组胃癌细胞中CD44 V6表达水平明显低于空白对照组。结论 :消痰散结方抑制胃癌细胞转移的环节可能和影响黏附分子的表达 ,从而影响细胞的黏附性机制有关     

Specific targeting of mitomycin C to tumors with the use of anti-gastric cancer monoclonal antibody

In the present study, an anti-gastric cancer monoclonal antibody, MGb2, was chosen to prepare antibody-mitomycin C (MMC) conjugate with dextran T-40 as intermediary. 20 molecules of MMC were introduced into each molecule of antibody, while the antigen-binding capacity of the antibody was kept well. The conjugate showed highly selective cytotoxicity upon human gastric cancer cell line SGC-7901. Radioimmunoimaging and biodistribution studies indicated that after conjugation with MMC via dextran T-40 as intermediary, the tumor localization capacity of the antibody was well retained. When tested in nude mice, by inoculation with human gastric carcinoma SGC-7901 in bilateral subrenal capsules, intraperitoneal injection of the conjugate daily for 6 days at the dose of 1 mg/kg of drug gave a tumor inhibitory rate of 68.67%, the result being far better than that of free MMC or irrelevant conjugate. No synergic effect was found in regard to the mixture of MGb2 with MMC.     

Specific targeting of mitomycin C to tumors by anti-gastric cancer monoclonal antibody.

In the present study, an anti-gastric cancer monoclonal antibody, MGb2, was chosen to prepare antibody-mitomycin C (MMC) conjugate with dextran T-40 as intermediary. Twenty molecules of MMC were introduced into each molecule of antibody while the antigen-binding capacity of the antibody was kept well. The conjugate showed selective cytotoxicity upon human gastric cancer cell line SGC-7901. Radioimmunoimaging and biodistribution studies indicated that after conjugation with MMC via dextran T-40 as intermediary, the tumor localization capacity of the antibody was well retained. When tested in nude mice, inoculated with human gastric carcinoma SGC-7901 in bilateral subrenal capsules, intraperitoneal injection of the conjugate daily for 6 days at a dose of 1 mg/kg gave a tumor inhibitory rate of 68.67%, which was far better than that of free MMC or irrelevant conjugate. No synergetic effect was found in regard to the mixture of MGb2 with MMC.

     

~(125)I-MGB_2-DM在荷胃癌裸鼠体内的定位

作者用碘~(125)标记由顺-乌头酸酐法制备的胃癌单抗(MGB_2)-柔红霉素(DM)结合物,观察其在荷人胃癌SGC-7901裸鼠体内分布状态.在MGB_2—DM中,每个抗体分子引入4～5个分子DM,抗体活性保存良好.结果表明,~(125)Ⅰ—MGB_2—DM能较满意地定位于肿瘤组织,腹腔给予~(125)Ⅰ-MGB_2 DM 96h,瘤/血比值达203,其结果与未修饰MGB_2相近.     

Tumor localization of anti-gastric cancer monoclonal antibody MGb_2-daunomycin conjugate in nude mice bearing human gastric carcinoma SGC-7901

In the present study,an anti-gastric cancer monoclonal antibody MGb_2-daunomycin(DM)conjugate was prepared by cis-aconitic anhydride method.The distribution of the conjugatein nude mice bearing human gastric carcinoma SGC-7901 was investigated.Four to five moleculesof DM were introduced into each molecule of antibody while the antigen-binding capacity ofthe antibody was well-retained.This conjugate could be satisfactorily localized in the tumortissue.Ninety-six hours after intraperitoneal injection of ~(125)I-MGb_2-DM conjugate,the ratio ofradioactivity per milligram of tumor to that per milligram of blood reached 203,the result beingquite similar to that of unmodified MGb_2.