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杂交瘤细胞株

杂交瘤细胞株的相关文献在1989年到2023年内共计1391篇,主要集中在畜牧、动物医学、狩猎、蚕、蜂、基础医学、内科学 等领域,其中期刊论文112篇、会议论文14篇、专利文献138081篇;相关期刊84种,包括生物技术通报、中国学术期刊文摘、中国免疫学杂志等; 相关会议12种,包括中国畜牧兽医学会家畜传染病分会第八次全国会员代表大会暨第15次学术研讨会、2012年中华医学会全国临床微生物学术交流大会、中国畜牧兽医学会2011学术年会等;杂交瘤细胞株的相关文献由2324位作者贡献,包括胥传来、刘丽强、匡华等。

杂交瘤细胞株—发文量

期刊论文>

论文:112 占比:0.08%

会议论文>

论文:14 占比:0.01%

专利文献>

论文:138081 占比:99.91%

总计:138207篇

杂交瘤细胞株—发文趋势图

杂交瘤细胞株

-研究学者

  • 胥传来
  • 刘丽强
  • 匡华
  • 宋珊珊
  • 徐丽广
  • 吴晓玲
  • 胡拥明
  • 马伟
  • 孙茂忠
  • 郝昌龙
  • 期刊论文
  • 会议论文
  • 专利文献

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    • 白云鹏; 曾玲; 徐彬瑗; 申海燕; 郝育杰; 周洋
    • 摘要: 目的 制备抗血管紧张素Ⅰ(angiotensin I,AⅠ)的杂交瘤细胞株,比较不同连接载体产出杂交瘤细胞的效果.方法 通过对载体分别为牛血清白蛋白(BSA)和卵清蛋白(OVA)的AⅠ-BSA和AⅠ-OVA免疫原进行小鼠免疫,将骨髓瘤细胞与免疫合格的小鼠进行脾细胞融合,筛选出阳性杂交瘤细胞株,经过克隆、扩大培养、腹水诱生等工序,生产出AⅠ单克隆抗体,测定并比较不同连接载体产出杂交瘤细胞抗体的性能,筛选具有应用价值的抗体.结果 本方法共产出9株AⅠ杂交瘤细胞株,其中AⅠ-OVA组产出4株,AⅠ-BSA组产出5株,进行各细胞抗体的功能检验,测定亚型、亲合常数,与AⅡ、1~7氨基酸肽段、5~8氨基酸肽段以及1~9氨基酸肽段均无交叉反应,优选出AⅠ-BSA组的D11C12单抗建立检测方法进行性能评价,空白限为0.04ng/mL,低值样本的分析内精密度为4.5%,高值样本的分析精密度为3.5%,与对照试剂盒进行68例样本检测结果相关性对比,r为0.9728.结论 成功建立抗血管紧张素Ⅰ杂交瘤细胞株制备方法,可以满足AⅠ单克隆抗体的使用要求.
    • 刘运超; 杨苏珍; 陈玉梅; 王聚财; 尚延丽; 魏蔷; 陈维聪; 冯华; 张改平
    • 摘要: 为了制备具有中和活性的抗猪细小病毒(Porcine parvovirus,PPV)单克隆抗体,采用血凝试验(HA)鉴定纯化的重组PPV VP2蛋白的活性,将重组VP2蛋白与弗氏佐剂混合后免疫BALB/c小鼠.免疫3次后,小鼠血清的血凝抑制(HI)效价可达1∶216,取免疫小鼠脾细胞与SP2/0细胞融合制备杂交瘤细胞,采用有限稀释法进行多轮亚克隆,经免疫过氧化物酶单层细胞试验(IPMA)筛选,成功获得杂交瘤细胞株5F7和11B3,能够稳定分泌中和性单克隆抗体.单克隆抗体5F7和11B3的轻链型均为Kappa,重链型分别为IgG2a和IgG2b.经ELISA和IPMA检测,单克隆抗体5F7和11B3均能与重组PPV VP2蛋白和PPV病毒粒子发生特异反应,而与猪圆环病毒2型(PCV2)、猪繁殖与呼吸综合征病毒(PRRSV)和猪瘟病毒(CSFV)无交叉反应.5F7和11B3腹水针对PPV病毒反应的ELSIA效价分别为1∶10240和1∶20480;针对PPV感染PK15细胞的中和效价分别为1∶211和1∶210.Western blot鉴定结果显示,单克隆抗体5F7和11B3均不与变性的VP2蛋白发生反应,说明2株单克隆抗体均识别重组PPV VP2蛋白的构象型表位.综上,成功制备了2株具有中和活性的抗PPV单克隆抗体.
    • 李翠娜; 王增利; 李春生; 董维亚; 韩庆安; 杜顺丰; 武孝丽; 吴萌
    • 摘要: 为了制备抗猪血红蛋白的单克隆抗体,试验用猪血红蛋白纯品为抗原,直接免疫雌性BALB/c小鼠;待血清检测为阳性后,将小鼠骨髓瘤细胞SP2/0与免疫后小鼠脾细胞融合;用间接ELISA法和有限稀释法进行阳性单克隆杂交瘤细胞株的筛选,并对其中效价较高的一株杂交瘤细胞株进行特性分析.采用辛酸-硫酸铵方法纯化抗体,用间接ELISA法测定亲和力常数.结果表明:该方法能得到稳定分泌抗猪血红蛋白单克隆抗体的杂交瘤细胞株并制备腹水,获得了效价为1:800 000的抗猪血红蛋白单克隆抗体,该单克隆抗体的亚型为lgG1,亲和力常数为2.8×108 L/mol,且该单克隆抗体与牛、人血红蛋白交差反应率均小于0.1%.说明该单克隆抗体特性良好,可以作为后续试验原料,进一步用于猪血红蛋白检测试剂盒的研制中.
    • 李延芳; 赵晴晴
    • 摘要: 杂交瘤细胞分泌单克隆抗体,在制备单克隆抗体的过程中,尤为重要的环节是筛选分泌特异性单克隆抗体的杂交瘤细胞株。脾细胞和骨髓瘤细胞2个细胞融合而成的细胞是杂交瘤细胞,基因表达不稳定,培养传代或冻存过程中,轻易会缺失或大幅度使分泌单克隆抗体的能力下降,所以在筛选出特异性杂交瘤细胞株后的一段时间内,要多次检测其分泌抗体的能力。一般实验室研究通过杂交瘤细胞技术,前期筛选出一株能分泌猪PD-L1蛋白单克隆抗体的杂交瘤细胞株3B5,利用酶联免疫吸附实验(enzyme linked immunodeficient assay,ELISA)来测定杂交瘤细胞3B5分泌单克隆抗体的上清效价,来研究杂交瘤细胞的稳定性,杂交瘤细胞株3B5在冻存后30天、60天和90天复苏,ELISA检测细胞培养上清的抗体效价。
    • 舒曼; 鲁卫东; 钱雯; 马波; 叶红艳; 黄微; 兰萍
    • 摘要: 目的:筛选能分泌A群脑膜炎奈瑟球菌多糖抗体的杂交瘤细胞株,制备单克隆抗体,运用该单抗建立检测ACYW135群脑膜炎奈瑟球菌多糖疫苗中A群多糖的竞争ELISA方法.方法:运用经典的杂交瘤技术筛选杂交瘤细胞株,采用小鼠腹腔注射收获腹水型单克隆抗体,优化条件建立间接竞争ELISA,分别测定5支同一批和3个不同批次的ACYW135群脑膜炎奈瑟球菌多糖疫苗成品中的A群多糖含量,检测结果应用SPSS软件做单样本t检验分析.结果:标准曲线经Log-logit处理,得到回归方程:y=-1.664 8-2.254 2x,R2=0.989 5,线性范围0.022 8~1.200 7μg/mL,IC50为0.165 8μg/mL,最低检测限为0.022 8 μg/mL,检测疫苗中A群多糖含量,检测值与疫苗标示量无统计学差异(P>0.05).结论:成功筛选杂交瘤细胞株,制备A群多糖特异性单克隆抗体,建立的间接竞争ELISA方法,灵敏度高、稳定性好、可重复性强,可用于脑膜炎奈瑟球菌多糖疫苗研发过程中的定量检测,能克服磷含量法和火箭免疫电泳法检测A群多糖含量的缺点.
    • 李新朋; 姜金庆; 钱爱东; 王自良; 范国英; 单晓峰; 康元环; 李艺
    • 摘要: 【目的】氟喹诺酮类药物(FQs)在畜牧业中广泛用于预防和治疗细菌性疾病,不合理使用导致在动物性食品中残留,严重危害消费者健康,其检测受到世界各国重视。制备抗多种FQs单克隆抗体(mAbs),建立间接竞争 ELISA(icELISA)检测方法,为动物性食品中 FQs 多残留检测奠定基础。【方法】环丙沙星(CPFX)羧基上引入氨基丁酸后为半抗原(CPFX-A),质谱鉴定;分别用碳二亚胺法(DDC)和混合酸干法将半抗原与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联合成免疫原(CPFX-A-BSA)和包被原(CPFX-A-OVA),紫外和红外光谱鉴定是否偶联成功;CPFX-A-BSA 免疫Balb/c小鼠,间接ELISA和icELISA方法测定多抗血清效价和敏感性,选取抗血清效价高和敏感性好的鼠用于细胞融合;NS0细胞和脾细胞按1﹕5比例在PEG-1500作用下融合,筛选抗FQs杂交瘤细胞株,并进行效价、亚型、敏感性和交叉反应率鉴定;体内诱生腹水法制备 mAb,液体石蜡作为诱导剂,每只鼠注射108个杂交瘤细胞;选敏感性和广谱特异性好的腹水mAb,建立icELISA标准曲线,对icELISA检测方法优化,在鸡肉中添加10种FQs进行测定,将测定结果与HPLC法比较,用SPSS 17.0软件进行显著差异性分析。【结果】半抗原改造和人工抗原合成成功;3只鼠血清效价均达到1﹕1.28×104以上,2号鼠血清效价最高(1﹕2.56×104),且IC50值最低(12.92 ng·mL-1),选择2号鼠作为细胞融合用鼠;4次亚克隆筛选并鉴定后获得3株敏感广谱特异的杂交瘤细胞,分别命名为2H5、3D11、4F4,细胞上清效价分别为1﹕1600、1﹕1600和1﹕800,腹水效价分别为1﹕1.6×106、1﹕8.2×105和1﹕8.2×105;icELISA方法的包被原浓度为1μg·mL-1,5%猪阴性血清4°C包被过夜,单抗1﹕40000稀释,山羊抗鼠酶标二抗(GaMIgG-HRP)1﹕8000稀释;反应温度均为37°C,标准品与单抗同时加入反应15 min,洗板后加二抗反应25 min;显色10 min,2H5细胞株腹水敏感性和广谱特异性最好,2H5的线性回归方程为y=-28.022x+56.219,R2=0.9782,对10种FQs的IC50值分别为环丙沙星(CPFX)1.67 ng·mL-1、诺氟沙星(NOR)1.82 ng·mL-1、培氟沙星(PEF)1.97 ng·mL-1、恩诺沙星(ENR)1.54 ng·mL-1、单诺沙星(DAN)2.79 ng·mL-1、洛美沙星(LOM)3.38 ng·mL-1、氧氟沙星(OFL)5.50 ng·mL-1、麻保沙星(MAR)4.40 ng·mL-1、沙拉沙星(SAR)11.76 ng·mL-1、二氟沙星(DIF)13.60 ng·mL-1,与10种FQs的交叉反应率为12.3%—108.4%,与非 FQs交叉反应率均低于0.01%,对10种FQs的检测限(LODs)为0.09 ng·mL-1—0.64 ng·mL-1。icELISA法鸡肉中10种FQs的回收率为80.5%—91.8%,HPLC法的回收率为85.1%—95.7%,二者变异系数均低于10.0%;两种检测方法差异不显著(P>0.05)。【结论】该试验获得了高效价、敏感、广谱特异性的mAbs,建立的icELISA方法可用于同时检测鸡肉中10种FQs。%Objective]Fluoroquinolones(FQs) are widely used in veterinary medicine for the treatment and prevention of bacterial infection. With the increasing use, FQs residues in animal edible tissues have caused serious public health problems and attracted serious attention by research scholars all over the world. The objective of this study was to produce class-specific monoclonal antibodies (mAbs) against fluoroquinolones (FQs), establish competitive indirect enzyme linked immunesorbent assay (icELISA), and in order to lay a foundation for detection of multi-residue FQs in animal foods.[Method]The aminobutyric acid was introduced to carboxyl of ciprofloxacin as hapten (CPFX-A) and was proved by (+) ESI-MS spectrum, which was conjugated to bovine serum albumin (BSA) as immunogen (CPFX-A-BSA) by the N,N'-Dicyclohexylcarbodiimide (DDC) method, and to ovalbumin (OVA) as coating antigen(CPFX-A-OVA) by mixed anhydride method, respectively, which were then identified by infrared ray (IR) and ultraviolet (UV). Balb/c mice immunized by CPFX-A-BSA were selected for cell fusion, which was identified by ELISA and icELISA. Under the effect of PEG-1500, NS0 cells and spleen cells were fused at the ratio of 1﹕5. Hybridoma lines that secrete mAb against FQs were selected and their immunological traits were characterized by titer, subtype, sensitivity and cross reaction rate, which ascites were carried out by injecting 108 hybridoma cells in vivo, and icELISA standard curve was established and optimized. High titer, class-specific monoclonal antibody was used to detect 10 FQs in chicken for calculating recovery rate and variation coefficient. The data were also compared with that of HPLC, and SPSS 17.0 software was used to conduct the significant difference analysis.[Result]The hapten and artificial antigen were synthesized successfully and antiserum titers of three mice were higher than 1﹕1.28×104, in which the titer and IC50 of No.2 mouse were 1﹕2.56×104 and 12.92 ng·mL-1. Three hybridoma cell lines named 2H5, 3D11, and 4F4 were screened after 4 times subclone, which titers were 1﹕1 600, 1﹕1 600, and 1﹕800 in supernatants and 1﹕1.6×106, 1﹕8.2×105, and 1﹕8.2×105 in ascites, respectively. The icELISA procedure was optimized at a concentration of CPFX-A-OVA for 1μg·mL-1 at 4°Cpackage overnight by 5% negative serum of pig, monoclonal antibody and GaM IgG-HRP were diluted 1﹕40 000 and 1﹕8 000, respectively. Under the reaction temperature of °C37, reaction time of standard substance and monoclonal antibody was 15 min, and 25 min after adding GaMIgG-HRP, also 10 min for termination reaction. Cell line named 2H5 showed a good sensitivity and class-specific toward 10 FQs, the linear regression equation was y=-28.022x+56.219,R2=0.9 782, with an IC50 value of 1.67 ng·mL-1 for ciprofloxacin, 1.82 ng·mL-1 for norfloxacin, 1.97 ng·mL-1 for pefloxacin, 1.54 ng·mL-1 for enrofloxacin, 2.79 ng·mL-1 for danofloxacin, 3.38 ng·mL-1 for lomefloxacin, 5.50 ng·mL-1 for ofloxacin, 4.40 ng·mL-1 for marbofloxacin, 11.76 ng·mL-1 for sarafloxacin, 13.60 ng·mL-1 for difloxacin, and the lowest detectable limits (LODs) of 0.09 ng·mL-1-0.64 ng·mL-1 and cross-reactivity (CR) of 12.3%-108.4%, no cross-reactivity to other compounds was found. The recovery ranges of 10 FQs spiked in chicken using icELISA were 80.5%-91.8%, 85.1%-95.7%for HPLC, both of the coefficient variations (CVs) were below 10.0%, and no significant difference (P>0.05) was observed.[Conclusion]The high-sensitivity and class-specific mAb against FQs was prepared, which laid a solid foundation for FQs multi-residue detection.
    • 胥亚; 许国莹; 周红; 解鸿翔; 夏龙飞; 刘敬敬; 张晓蕾
    • 摘要: Objective:To produce and identify the monoclonal antibodies (MAbs) against β2-glycoprotein Ⅰ (β2 GP Ⅰ).Methods:The spleen cells from the BALB/c female mouse immunized with β2 GP Ⅰ were fused with SP2/0 mouse myeloma cells in PEG to generate hybridoma cells,which were screened by limited dilution.Then the hybridoma cells were injected to BALB/c mice to make ascites,in which the antibodies were produced abundantly,followed by purified with ammonium sulfate and protein G.Colchicine was used to identify the number of chromosome of hybridoma cells.The properties of the MAbs against β2 GP Ⅰ (the titer of the MAbs in the culture medium and ascites,the isotypes of the MAbs and the specificity of the MAbs) were determined by indirect ELISA,MAbs isotyping kit and Western blot,respectively.Results:One hybridoma cell line (β2),secreting MAbs against β2 GP Ⅰ,had been established.The number of chromosome of the hybridoma cells was between 99 and 108.Further studies found that the titer of the MAbs against β2GP Ⅰ in the culture medium and ascites was 1∶29 and 1∶215,respectively.The immunoglobulin of the MAbs was classified as IgG1/κ,with satisfactory specificity.Conclusion:The MAbs against β2GP Ⅰ were prepared and identified,which pave the way for the research in the mechanism of antiphospholipid syndrome.%目的:制备β2糖蛋白Ⅰ(β2GPⅠ)单克隆抗体(MAbs),并鉴定抗体的特性.方法:以纯化的人血浆β2GP Ⅰ免疫BALB/c小鼠,采用PEG融合技术,间接ELISA法和有限稀释法,制备和筛选杂交瘤细胞.杂交瘤细胞继续扩大培养后注入BALB/c小鼠腹腔制备腹水,经硫酸铵沉淀及Protein G纯化MAbs.秋水仙素阻抑法鉴定杂交瘤细胞株染色体数目,ELISA测定滴度,Ig亚类试剂盒鉴定Ig亚类,Western blot鉴定特异性.结果:获得1株稳定分泌MAbs的杂交瘤细胞株β2,该杂交瘤细胞株的染色体数为99 ~108.进一步研究发现,其培养液和腹水滴度为1∶29和1∶215,其MAbs分类为IgG1/κ,Western blot 显示纯化的MAbs及标准anti-β2GP Ⅰ与β2GP Ⅰ均有反应条带,具有较好的特异性.结论:成功获得并纯化β2GP Ⅰ单克隆抗体,为进一步研究β2GP Ⅰ/anti-β2GP Ⅰ复合物在抗磷脂综合征中的作用奠定了基础.
    • 乔元彪; 王权; 张文举; 陈沁
    • 摘要: 目的:制备特异性强、灵敏度高的人促甲状腺激素(hTSH)单克隆抗体,为建立高灵敏性和高特异性的hTSH检测方法奠定基础.方法:人工合成人促甲状腺激素(hTSH)上的28个氨基酸,采用戊二醛交联法与牛血清白蛋白(BSA)构建完全抗原.用人工免疫原免疫BALB/c小鼠,利用杂交瘤技术建立分泌抗hTSH单克隆抗体的杂交瘤细胞,并对得到的单抗进行鉴定.结果:建立了一株杂交瘤细胞4E5,制备了腹水单抗,经鉴定,抗体重链属于IgG1类型,轻链属于κ型,McAb腹水ELISA效价达1:1.6×105,与促黄体生成激素(LH)、促卵泡激素(FSH)、人绒毛膜促性腺激素(HCG)无明显交叉反应,细胞株冻存复苏后抗体分泌稳定.结论:本实验建立的细胞株显示稳定性好,特异性高,McAb腹水效价高,为进一步提高hTSH检测的灵敏性和简便性提供依据.
    • 张越; 屈会化; 吴婷婷; 薛瑾; 孙晔; 孙慧; 李翼飞; 赵琰; 王庆国
    • 摘要: Objective:To prepare a specific monoclonal antibody against glycyrrhizic acid (GA).Methods:BALB/c mice were immunized with the prepared GA-BSA conjugate.When the serum detection was positive,spleen cells from the mice were isolated and fused with SP2/0 myeloma cells at a ratio of 10∶ 1.Monoclonal hybridoma cells were screened by indirect ELISA and limited dilution.The ascites antibodies were induced and prepared by the positive cell line,and then were purified by caprylic acid/ammonium sulfate precipitation and tested by cross reaction.Results:The monoclonal antibody hybridoma cell line GA-Mab-DF5 steadily secreting glycyrrhizic acid antibodies was obtained,the purity of ascites antibodies was up to 98%,and the recovery rate was 80%.The linear range was 4 μg · mL-1 to 64 μg · mL-1 (R2 =0.9986) and no cross-reactivity was detected with BSA,gelatin,cholic acid,deoxycholic acid,paeoniflorin,baicalin and so on.Conclusion:The anti-GA antibody-secreting hybridomas are obtained with high sensitivity and specificity,which lays a foundation for the trace detection and immunoaffinity column preparation of glycyrrhizic acid in samples.%目的:制备特异性的甘草酸(GA)单克隆抗体.方法:用合成的甘草酸-牛血清白蛋白(GA-BSA)人工抗原免疫BALB/c小鼠;待血清检测阳性后,取其脾细胞与小鼠骨髓瘤细胞SP2/0按10∶1的比例融合;用间接竞争ELISA法和有限稀释法进行单克隆杂交瘤细胞的筛选;用阳性细胞株诱导制备腹水抗体;用辛酸硫酸铵法纯化抗体,并进行交叉反应检测.结果:获得了稳定分泌抗甘草酸的单克隆抗体杂交瘤细胞株GA-Mab-DF5,腹水抗体纯化后纯度达98%,回收率达80%.线性检测范围为4~ 64 μg·mL-1,R2 =0.9986,并且与BSA、明胶、胆酸、去氧胆酸、芍药苷、黄芩苷等无交叉反应.结论:成功获得能稳定分泌甘草酸单克隆抗体的杂交瘤细胞株,灵敏度、特异性均较高,为样品中甘草酸的快速微量检测及免疫亲和色谱柱制备奠定了基础.
    • 孙婧; 毕振威; 夏兴霞; 王晓丽; 诸玉梅; 董晨红; 贾红; 朱瑞良; 王永山
    • 摘要: 为建立方便快捷的犬瘟热病毒(CDV)检测方法,本实验应用杂交瘤细胞融合技术,建立了4株分泌抗CDV单克隆抗体(MAb)的杂交瘤细胞株,分别命名为A2、F7、H4和G12.相加ELISA试验显示4株MAb作用于CDV不同的抗原表位.选取相加指数较高的两株MAb G12和A2分别作为捕获抗体和酶标检测抗体,建立检测CDV的夹心ELISA检测方法,并对实验条件进行了优化.结果显示,该方法与犬细小病毒、犬副流感病毒、犬腺病毒1型、犬冠状病毒等犬类病毒无交叉反应,敏感度为5 μg/mL,变异系数小于6%.利用该方法与RT-PCR方法同时检测57份临床样品,两种方法的符合率为100%.本实验建立的MAb夹心ELISA方法具有特异、敏感、方便快捷等优点,适用于大批量临床样品检测.%To develop a method for the detection of canine distemper virus (CDV),four hybridoma of A2,F7,H4 and G12 secreting monoclonal antibodies (MAb) to CDV were prepared by the fusion of SP2/0 cells and splenocyte from BALB/C mice immunized with the purified CDV.Additivity ELISA revealed that the four MAbs recognized spatially independent epitopes of CDV.In addition,a sandwich ELISA was established with MAb G12 as capture antibody and HRP conjugated MAb A2 as detection antibody for the rapid detection of CDV which had no cross-reactivity with other canine virus and the detection limit was 5 μg/mL.The coefficient of variation of reproducibility was less than 6%.A total of 57 clinical samples were detected by the ELISA and RT-PCR with agreement rate of 100%.The result indicated that the sandwich ELISA was applicable to the clinical detection of CDV.
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