【正文】 . at room temp. Then gently wiggle the tube for 37176。 Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies ? Production of monoclonal antibodies involves in vivo or in vitro procedures or binations thereof. Before production of antibodies by either method, hybrid cells that will produce the antibodies are generated. The steps in producing those cells are outlined below。 3－ 4天后，在無菌條件下取出脾或淋巴結(jié)制成懸液，存活率在 95％以上的可以用于融合。 免疫方法 ：體內(nèi)法或體外法。多用 BALB/C 小鼠的骨髓瘤細(xì)胞。 antibodies produced by conventional methods, on the other hand, are derived from preparations containing many kinds of cells, and hence are called polyclonal. An example of how monoclonal antibodies are derived is described below ? A myeloma is a tumor of the bone marrow that can be adapted to grow permanently in cell culture. When myeloma cells were fused with antibodyproducing mammalian spleen cells, it was found that the resulting hybrid cells, or hybridomas, produced large amounts of monoclonal antibody. This product of cell fusion bined the desired qualities of the two different types of cells: the ability to grow continually, and the ability to produce large amounts of pure antibody. ? Because selected hybrid cells produce only one specific antibody, they are more pure than the polyclonal antibodies produced by conventional techniques. They are potentially more effective than conventional drugs in fighting disease, since drugs attack not only the foreign substance but the body39。 單抗的制備過程 ? 動物免疫與親本細(xì)胞的選擇 ? 細(xì)胞融合：淋巴細(xì)胞雜交瘤的制備 ? 雜交瘤細(xì)胞的篩選：有限稀釋法等 ? 單克隆抗體的制備和凍存 ? 單克隆抗體的純化 ? Monoclonal antibody technology allows us to produce large amounts of pure antibodies in the following way: We can obtain cells that produce antibodies naturally。雜交瘤細(xì)胞既能在體外無限增殖，又能持續(xù)分泌成分單一的特異性抗體。 classic examples are the antibodies to the childhood diseases chickenpox and measles. ? The second characteristic of antibodies makes it possible to develop vaccines. A vaccine is a preparation of killed or weakened bacteria or viruses that, when introduced into the body, stimulates the production of antibodies against the antigens it contains. ? It is the first trait of antibodies, their specificity, that makes monoclonal antibody technology so valuable. Not only can antibodies be used therapeutically, to protect against disease。 類別，表位，個體間比例，時間 Antibodies that arise in an animal in response to typical antigens are heterogenerous, because they are formed by several different clones of plasma cellsPolyclonal. Antibodies that arise from a single clone of cells, eg, in plasma cell tumor(myeloma) are homogeneous, they are monoclonal. Antibodies are globulin proteins that react specifically with the antigen that stimulated their production. There are five classes of antibodies based on differences on their heavy chains. Functions Neutralize toxins and virus Opsonize microbes Activate plements Prevent attachment Catalytic capability ? Monoclonal Antibody Technology The Basics ? Substances foreign to the body, such as diseasecausing bacteria and viruses and other infectious agents, known as antigens, are recognized by the body39。s immune system as invaders. 動物脾臟有上百萬個 B淋巴 細(xì)胞，一個細(xì)胞就是一個克隆，它針對抗原上的一個抗原決定 簇 產(chǎn)生的抗體是單克隆抗體。特點(diǎn)：外源性、結(jié)構(gòu)性（分子表面具有穩(wěn)定的環(huán)狀結(jié)構(gòu)基團(tuán)作為識別位點(diǎn)）、特異性（抗原與抗體的反應(yīng)）。第四章 2 單克隆抗體 Monoclonal Antibodies 單克隆抗體 抗體的結(jié)構(gòu)與功能 單克隆抗體的制備和生產(chǎn) 單克隆抗體的改造和應(yīng)用 Human lymphoid ans ? Central lymphoid ans?? ? Secondary lymphoid ans? ? Thymus, bone marrow ? Tonsil, adenoid, lymphatic vessels, lymph nodes, spleen , peyer’s patches in small intestine, appendix ? Innate immune response ? Adaptive immune response Humoral immunity Cellmediated immunity 抗原： 進(jìn)入機(jī)體內(nèi)對機(jī)體的免疫系統(tǒng)產(chǎn)生刺激作用的外源物質(zhì)。包括：蛋白質(zhì)、多糖、核酸、病毒、細(xì)菌、各種細(xì)胞等。 Antigens are molecules that react with antibodies, whereas immunogens are molecules that induce an immune responses. Substances foreign to the body, such