共查询到19条相似文献,搜索用时 140 毫秒
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目的:研制抗人喉癌/抗血管内皮因子(VEGF)双功能克隆抗体,用于喉癌抗血管生成治疗。方法:采用二次杂交瘤技术制备抗人喉癌/抗VEGF双功能抗体。经酶联免疫吸附试验法和SP法检测喉癌及癌前病患者血清及癌组织中VEGF的含量表达。结果:获得6株分泌抗人喉癌/抗VEGF双功能抗体的杂交瘤,经免疫组化证实与喉癌细胞特异性结合率为93%,而与血管内皮细胞结合率为89%。血清中VEGF含量表达,喉癌组与癌前病组及正常对照组相比差异均显著。IgG亚型鉴定为IgG2aBSAb抗体效价为1:25 600倍(ELISA法)。结论:二次杂交瘤法制备的双功能抗体具有均匀性、可控性、效价高、稳定性好,可用于喉癌抗血管生成治疗,动态检测可作为判断喉癌预后的客观指标。 相似文献
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人胃癌双功能抗体制备的研究 总被引:1,自引:0,他引:1
以不同技术路线制备P210/MMC,P210/TNP及P40/CD3三种双功能抗体(BsAb)。在11次细胞融合中,接种7484孔,获15株二次杂交瘤,其中5株分泌BsAb。将亲本杂交瘤分别诱变为HGPRT^-及TK^-,融合后以HAT选择培养并辅以饲养细胞较易获得二次杂交瘤。证明两种抗原交互反应的桥联试验,特别是当二次杂交瘤分泌不同类别的Ig时,对于确证BsAb的存在有着重要意义。本文首次报告, 相似文献
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目的:构建并表达抗人精浆蛋白/抗CD3的双特异性单链抗体(BsscFv),并检测其生物学活性。方法:利用重叠延伸拼接PCR,拼接抗人精浆蛋白scFv基因和抗CD3scFv基因,并在中间引入柔性短肽(GlySerGly)2,构建抗人精浆蛋白/抗CD3的BsscFv基因。测序正确后,将融合基因亚克隆入真核表达载体中,并在HeLa细胞中进行表达,采用流式细胞术(FCM)和51Cr释放试验,评价BsscFv的抗原结合活性和体外介导的特异性杀伤靶细胞的效应,以及利用裸鼠前列腺癌模型观察其在体内的抑瘤作用。结果:测序分析证实,Bss-cFv基因片段的大小为1.5kb,编码500个氨基酸,该序列与设计的完全一致。SDS-PAGE和Westernblot分析证明:表达产物存在于HeLa细胞的培养上清中,其相对分子质量(Mr)为61000。FCM结果显示:BsscFv可特异性的结合前列腺癌细胞LNCaP和CD3 淋巴瘤细胞Jurkat,结合率分别为54.1%和53.7%。体外实验表明,BsscFv可介导CTL对LNCaP细胞的杀伤。与对照组相比较,接种LNCaP的裸鼠在体内注射激活的CTL和BsscFv治疗后,肿瘤的生长明显受到抑制(P<0.05)。结论:抗人精浆蛋白/抗CD3的BsscFv具有一定的生物学活性,在体内、体外均可介导CTL杀伤靶细胞LNCaP。 相似文献
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微型双功能抗体抗CD3/抗CD20的构建和表达 总被引:2,自引:2,他引:0
目的:构建和表达抗CD3/抗CD20微型双功能抗体,并测定微型双功能抗体的生物学活性。方法:采用PCR和overlap PCR方法构建抗CD3/抗CD20微型双功能抗体,并用双脱氧终止法测定DNA序列;采用亲和层析法纯化该产物,并用Western blot和分了排阻层析鉴定纯化产物;采用FACS法和玫瑰花环试验鉴定纯化产物与靶细胞的结合活性。结果:DNA序列测定结果表明:抗CD3/抗CD20微型双功能抗体已构建成功,表达可溶性产物的产量达1mg/ml以上,纯化产物中二聚体的比例达90%,具有与Jurkat(CD3^ 0和Daudi细胞(CD20^ )结合的活性,且能同时与Jurkat和Daudi细胞结合形成玫瑰花环。结论:利用Diabody形式,首次成功地构建了抗CD3/抗CD20微型双功能抗体,并获得较高表达,表达产物具有与相应2个靶抗原结合的活性。 相似文献
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在用纯HBsAg免疫山羊而产生的抗血清中,发现存在一种既能同HBsAg反应又能同时同正常人血清中某一成分起反应的微量的特殊抗体,验证试验的结果,证实了这一特殊抗体是一种双功能抗体,同这种功能抗体起反主尖的正常人血清成分可能是α1-抗胰蛋白酶。 相似文献
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双特异性抗体对LAK细胞增殖和细胞毒作用影响的体外研究 总被引:3,自引:0,他引:3
将抗CD3与抗HBs的单克隆抗体经化学偶联得到双特异性抗体,观察该双特异性抗体对LAK细胞增殖和增强细胞毒性的作用。结果显示双特异性抗体显著提高LAK细胞与2.2.15细胞结合率;促进淋巴细胞增殖。125I-UdR释放试验检测发现双特异性抗体增强LAK细胞对2.2.15细胞的细胞毒性且与抗体浓度呈正相关。进一步对抗体的特异性研究表明,加入双特异性抗体后LAK细胞对2.2.15细胞毒性作用显著高于对照组。 相似文献
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Maryam Balibegloo 《Expert Review of Clinical Immunology》2020,16(7):689-709
ABSTRACT
Introduction
Treatment of colorectal cancer as one of the most commonly diagnosed and a frequent cause of cancer-related deaths is of great challenges in health-related issues. 相似文献14.
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A recombinant bispecific single-chain antibody induces targeted,supra-agonistic CD28-stimulation and tumor cell killing 总被引:3,自引:0,他引:3
Grosse-Hovest L Hartlapp I Marwan W Brem G Rammensee HG Jung G 《European journal of immunology》2003,33(5):1334-1340
Endowing tumor cells with costimulatory signals for T cell activation has emerged as a promising strategy for tumor immunotherapy. Costimulatory molecules were either transfected into tumor cells to generate vaccines or were fused, e.g. to antibodies against tumor-associated antigens, to achieve targeted T cell costimulation in vivo. Here we report the production and purification of rM28, a recombinant bispecific single-chain antibody directed to a melanoma-associated proteoglycan and to the costimulatory CD28 molecule on human T cells. We found that a dimer of the recombinant molecule, bound to tumor target cells, induced pronounced T cell activation in peripheral blood mononuclear cell preparations without additional TCR/CD3 stimulation being required. The lytic activity generated after 3 days of stimulation effectively prevented tumor cell growth. However, it was unspecific and predominantly mediated by non T cells. Our findings demonstrate that presentation of a CD28 antibody within a suitable recombinant, bispecific format may result in a "targeted supra-agonistic stimulation" of the CD28 molecule, which leads to effective tumor cell killing after induction of unspecifically lytic cells. 相似文献
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目的 探讨用生物学方法治疗病毒感染性疾病及肿瘤的新途径。方法 以有重要补体活化调节功能的膜补体调节蛋白CD5 5为靶点 ,以 β GaL为模拟病毒或肿瘤抗原 ,制备“IgG”型抗CD5 5×抗 β Gal基因工程双特异性抗体 ,并对该重组抗体真核表达后的结合活性进行初步的验证。结果 克隆的CD5 5抗体可变区片段为新的小鼠抗体可变区片段 ,经HEK 2 93细胞表达后的重组抗体显示了良好的CD5 5及Fc结合活性。结论 本研究为病毒性疾病或肿瘤的免疫学治疗提供了新的途径及实验依据 相似文献
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用CDR3导向抗体库法对鼠抗人膀胱癌单抗进行人源化 总被引:1,自引:0,他引:1
目的 构建含鼠单抗CDR3的人源噬菌体抗体库 ,对膀胱癌鼠单抗进行人源化。方法通过重叠PCR及DNA重组技术 ,将抗膀胱癌鼠单抗BDI的CDR3区与人淋巴细胞来源的多样化的VH和Vκ组合 ,构建含BDICDR3的人源噬菌体抗体库 ;利用loxp cre定位重组系统 ,增加轻、重链的组合配对 ,获大容量抗体库 ;用膀胱癌细胞 (EJ)从中筛选人源化抗膀胱癌单抗Fab段 ,ELISA方法对所筛克隆进行特异性鉴定 ,同时运用竞争抑制实验对所筛特异性克隆进行抗原表位的核实。结果 首先构建了库容为 2× 10 7的含鼠CDR3区的人源噬菌体抗体库 ;经过在cre 细菌胞内的定位重组后 ,获得库容为 10 1 0 的大容量噬菌体抗体库 ;用EJ细胞筛选到可与其特异结合的人源性Fab段 ;竞争抑制实验表明所获人源抗体片段与亲本鼠源单抗结合同一抗原表位。结论 在没有亲本鼠单抗基因 ,仅了解其CDR3序列的条件下 ,可通过CDR3导向 ,构建大容量抗体库 ,筛选到相应的人源化抗体基因。 相似文献
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Generation and characterization of a novel tetravalent bispecific antibody that binds to hepatitis B virus surface antigens 总被引:5,自引:0,他引:5
Sung Sup Park Chun Jeih Ryu Young Jun Kang S. V. S. Kashmiri Hyo Jeong Hong 《Molecular immunology》2000,37(18):752-1130
Hepatitis B virus (HBV) infection is a worldwide public health problem affecting about 350 million people. HBV envelope contains three surface antigens, called pre-S1, pre-S2 and S. For the prophylaxis of HBV infection, only an anti-S monoclonal antibody was tested for the protective efficacy against HBV infection, but it was shown to be incomplete. In addition, some immune escape mutants carrying mutations on the S antigen were reported. Therefore, a multivalent bispecific antibody rather than a single monoclonal antibody would be more beneficial for the prophylaxis of HBV infection. We have generated a novel tetravalent bispecific antibody with two binding sites for each of the S and pre-S2 antigens. Each of the antigen-binding sites was composed of a single-chain Fv (ScFv). The tetravalent antibody was generated by constructing a single gene encoding a single-chain protein. This protein consisted of an anti-S ScFv whose carboxyl end was tethered, through a 45 amino acid linker, to the amino terminus of anti-preS2 ScFv that in turn was joined to the hinge region of human γ1 constant region. The single-chain protein was expressed in Chinese hamster ovary cells and secreted in culture supernatant as a homodimeric molecule. The tetravalent bispecific antibody showed both anti-S and anti-pre-S2 binding activities. In addition, the binding affinity of the bispecific antiboy for HBV particles was greater than that of either parental antibody. The tetravalent bispecific antibody is a potentially useful reagent for the prevention and treatment of HBV infection. 相似文献