血清中抗体的水平IgG初次免疫二次免疫igM40361257初次抗原注射后的周次抗原刺激的第一至三周，IgM生成；抗原再刺激后，IgG将取代IgM动物血清中产生抗体的时间示意图
动物血清中产生抗体的时间示意图 抗原刺激的第一至三周，IgM生成；抗原再 刺激后， IgＧ将取代IgM
抗体研究的发展阶段：①以1890年Behring发现白喉抗毒素为代表,用抗原免疫动物来获得多克隆抗体②以1975年Milstein&Kohler创建杂交瘤技术制备单克隆抗体(McAb为代表③以1994年Winter创建了噬菌体抗体库(repertoire of antibodies displayed onphages)技术,以基因工程方法制备抗体为代表。在此基础上发展成抗体工程
抗体研究的发展阶段： ①以1890年Behring发现白喉抗毒素为代表,用 抗原免疫动物来获得多克隆抗体。 ②以1975年Milstein & Kohler创建杂交瘤技 术制备单克隆抗体(McAb)为代表。 ③以1994年Winter,创建了噬菌体抗体库 (repertoire of antibodies displayed on phages)技术,以基因工程方法制备抗体为代 表。在此基础上发展成抗体工程
·多克隆抗体一种抗原具有多个抗原决定簇，每个抗原决定簇都能激一个B细胞产生一种抗体。这样所获得的免疫血清是多种抗体的混和物。·单克隆抗体(Monoclonal antibody McAb) :单一的B由一个抗原决定簇刺激的、细胞和骨髓瘤细胞融合增殖后所产生的、高度均一的抗体
• 多克隆抗体: 一种抗原具有多个抗原决定簇,每个 抗原决定簇都能刺激一个B细胞产生一种 抗体。这样所获得的免疫血清是多种抗 体的混和物。 • 单克隆抗体(Monoclonal antibody McAb): 由一个抗原决定簇刺激的、单一的B 细胞和骨髓瘤细胞融合增殖后所产生的、 高度均一的抗体。 3 1 2 4
多克隆抗体和单克隆抗体AdaptedfromMilstein(1980)ScientificAmerican,Oct.p.58AntigenSpleen cellsImmunizationB cellMyeloma23Cell fusion?MonoclonalPantibodiesY欧苗Antiseum伊76Pure sinaleAbIfasingleBcell waspickedupanacuitured,thenitwilproduce onlyonekindofantibodyAmixtureofallAbIf Bcell isfusedwithmyeloma,thefusedcellmightbeculturedand produceantibody.Eachhybridomalinecanproducepuresingleantibody,calledmonoclonalantibody
多克隆抗体和单克隆抗体 Pure single Ab After immunization, the mouse spleen contains B cells producing specific antibodies. Each B cell produces only one kind of antibody, which binds to i ts specific antigen. Conventional antiserum is the mixture of all antibodies produced by B cells from spleen. If a single B cell was picked up and cultured, then it will prod uce only one kind of antibody. But B cell can not survive well in the culture. Each hybridoma line can produce pure single antibody, called mon oclonal antibody. If B cell is fused with myeloma, the fused cell might be culture d and produce antibody. Adapted from Milstein (1980) Scientific American, Oct. p.58 1 2 3 4 m m m m 1 2 3 4 1 2 3 4 Monoclonal antibodies Cell fusion Spleen cells + Myeloma x Antiseum Antigen Immunization A mixture of all Ab 31 2 4 1 2 3 4 31 2 4 B cell Pure single Ab After immunization, the mouse spleen contains B cells producing specific antibodies. Each B cell produces only one kind of antibody, which binds to i ts specific antigen. Conventional antiserum is the mixture of all antibodies produced by B cells from spleen. If a single B cell was picked up and cultured, then it will prod uce only one kind of antibody. But B cell can not survive well in the culture. Each hybridoma line can produce pure single antibody, called mon oclonal antibody. If B cell is fused with myeloma, the fused cell might be culture d and produce antibody. Adapted from Milstein (1980) Scientific American, Oct. p.58 1 2 3 4 m m m m 1 2 3 4 1 2 3 4 Monoclonal antibodies Cell fusion Spleen cells + Myeloma x Antiseum Antigen Immunization A mixture of all Ab 31 2 4 1 2 3 4 31 2 4 B cell
二.单克隆抗体的制备McAb（Monoclonalantibody，单克隆抗体）：将在体外不能长期传代的抗体产生细胞（如Balb/C小鼠的免疫脾细胞）与在体外能迅速增殖并带有抗药标记的骨髓瘤细胞（如小鼠的NS-1或SP2/0）在聚乙二醇（PEG）作用下融合，然后利用抗药性标记和经稀释法选出单个产生抗体的杂交瘤细胞，再用血清学方法证实其与抗原结合的特异性和亲和力。这样的杂交瘤细胞能长期传代同时分泌针对单一抗原上单一抗原表位的抗体，称为McAb（第二代抗体）
McAb（Monoclonal antibody，单克隆抗体）： 将在体外不能长期传代的抗体产生细胞（如 Balb/C小鼠的免疫脾细胞）与在体外能迅速增殖并带 有抗药标记的骨髓瘤细胞（如小鼠的NS-1或SP2/0）在 聚乙二醇（PEG）作用下融合，然后利用抗药性标记 和经稀释法选出单个产生抗体的杂交瘤细胞，再用血 清学方法证实其与抗原结合的特异性和亲和力。这样 的杂交瘤细胞能长期传代同时分泌针对单一抗原上单 一抗原表位的抗体，称为McAb（第二代抗体）。 二. 单克隆抗体的制备