| 人血浆LDL的分离纯化及兔抗人apoB-100抗体的制备 |
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| 引用本文： | 沈瑶杰,徐亚雅,李芳芳,王杨萍.人血浆LDL的分离纯化及兔抗人apoB-100抗体的制备[J].生物磁学,2009(14):2652-2654,2666. |
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| 作者姓名： | 沈瑶杰 徐亚雅 李芳芳 王杨萍 |
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| 作者单位： | 浙江师范大学行知学院化学与生命科学学院,浙江金华321004 |
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| 摘 要： | 目的：为分离得到较高纯度的人血载脂蛋白B-100（apoB-100）及制备高效的兔抗人apoB-100抗体。方法：采用两步密度梯度离心分离人血浆得到LDL，产物经6％琼脂糖凝胶（Sepharose-6B）分子筛分离纯化后，再由透析浓缩获得纯化产物即apoB-100，同时用5％聚丙烯酰胺凝胶（PAGE）电泳鉴定为一条带。用分离纯化得到的apoB-100与完全及不完全福氏佐剂混合，以此作为免疫原经4-3-2—2-1（周）免疫新西兰大白兔，随后心脏取血制备兔抗人apoB-100抗体。结论：分离纯化apoB-100浓度为1．4625mg／ml，制备的兔抗体经免疫双扩散测定其效价为1：32。
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| 关 键 词： | apoB-100 密度梯度离心 纯化 免疫双扩散 抗体 |
Purification of LDL from Human Blood Plasma and Preparation of ApoB- 100 Antibody in Rabbit |
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| SHEN Yao-jie,XU Ya-ya,LI Fang-fang,WANG Yang-ping.Purification of LDL from Human Blood Plasma and Preparation of ApoB- 100 Antibody in Rabbit[J].Biomagnetism,2009(14):2652-2654,2666. |
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| Authors: | SHEN Yao-jie XU Ya-ya LI Fang-fang WANG Yang-ping |
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| Institution: | (Xingzhi College, College of Chemistry and Life Science, Zhejiang Normal University, Jinhua, Zhejiang 321004, China ) |
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| Abstract: | Objective： Apolipoprotein （apo） B-100 was extracted and purified from human blood plasma, and acquired the efficient antibody from the rabbit. Methods： LDL was extracted and purified using density gradient centrifuge, purified with 6% sepharose-6B gel filtration in this experiment. Then the purification was qualified by PAGE. The New Zealand rabbit was immunized by the apoB-100 with complete or incomplete Freud＇s adjuvant every 4-3-2-2-1 （week） injections. Subsequently, the antibody was acquired from the rabbit heart blood. Results： The concentration of apoB-100 reached 1.4625 mg/ml and its effect price evaluated by immunodiffusion method was 1 ： 32. |
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| Keywords: | ApoB-100 Density Gradient Centrifuge purification Immunodiffusion Antibody |
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