PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES TO THE EXTRACELLULAR DOMAIN OF PO
PRODUCTION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES TO THE EXTRACELLULAR DOMAIN OF PO
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DOI：
10.1002/jnr.490350107
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发表时间：
1993-05-01
影响因子：
4.2
通讯作者：
HARTUNG, HP
中科院分区：
文献类型：
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作者：
ARCHELOS, JJ;ROGGENBUCK, K;HARTUNG, HP
七个单克隆抗体提出了对免疫球蛋白样细胞外结构域（P0-艾德），周围神经系统髓鞘的主要蛋白质。用纯化的重组大鼠P0-ED免疫小鼠。融合后，通过ELISA筛选出7个识别重组、大鼠、小鼠或人P0-艾德的克隆（P01-P07），并通过Western blot、免疫组织化学和竞争试验对其进行表征。抗体属于IgG或IgM类，和P04-P07，与P0在新鲜冷冻和石蜡包埋切片的人或大鼠周围神经，但不与髓鞘蛋白的中枢神经系统的任何物种。通过竞争酶联免疫吸附试验（ELISA）和直接ELISA使用短合成肽跨越整个细胞外结构域的P0的抗体的表位特异性。结果表明，P01和P02在Western blot和免疫组化中表现出相同的反应模式，与P0的不同远距离表位反应。此外，单克隆抗体P05和P06识别P0的神经突原性细胞外序列内紧密接近的2个不同表位。这组单克隆抗体，每一个结合到P0的胞外结构域的不同表位，将是有用的，在体外和体内研究，旨在探索P0在髓鞘形成和脱髓鞘疾病的周围神经系统的作用。
Seven monoclonal antibodies were raised against the immunoglobulin-like extracellular domain of (P0-ED), the major protein of peripheral nervous system myelin. Mice were immunized with purified recombinant rat P0-ED. After fusion, 7 clones (P01-P07) recognizing either recombinant, rat, mouse, or human P0-ED were selected by ELISA and were characterized by Western blot, immunohistochemistry, and a competition assay. Antibodies belonged to the IgG or IgM class, and P04-P07, reacted with P0 in fresh-frozen and paraffin-embedded sections of human or rat peripheral nerve, but not with myelin proteins of the central nervous system of either species. Epitope specificity of the antibodies was determined by a competition enzyme-linked immunosorbent assay (ELISA) and a direct ELISA using short synthetic peptides spanning the entire extracellular domain of P0. These assays showed that P01 and P02 exhibiting the same reaction pattern in Western blot and immunohistochemistry reacted with different distant epitopes of P0. Furthermore, the monoclonal antibodies P05 and P06 recognized 2 different epitopes in close proximity within the neuritogenic extracellular sequence of P0. This panel of monoclonal antibodies, each binding to a different epitope of the extracellular domain of P0, will be useful for in vitro and in vivo studies designed to explore the role of P0 during myelination and in demyelinating diseases of the peripheral nervous system.