QUANTITATION OF BOVINE IMMUNOGLOBULIN ISOTYPES AND ALLOTYPES USING MONOCLONAL-ANTIBODIES
QUANTITATION OF BOVINE IMMUNOGLOBULIN ISOTYPES AND ALLOTYPES USING MONOCLONAL-ANTIBODIES
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DOI：
10.1016/0165-2427(90)90042-q
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发表时间：
1990-03-01
影响因子：
1.8
通讯作者：
NAESSENS, J
中科院分区：
文献类型：
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作者：
WILLIAMS, DJL;NEWSON, J;NAESSENS, J
制备了一组10种对牛免疫球蛋白M、A、G1、G2和轻链具有特异性的单克隆抗体，并开发了酶联免疫吸附试验，以使用该组单克隆抗体测量体液和培养上清液中的IG水平。抑制ELISA对于高亲和力的MAb是准确和灵敏的，使用高亲和力MAb，IL-A50（解离常数= 1.3 × 10 - 4）检测低至10 ng ml-1水平的IgM。10-11米）。对于亲和力较低的MAb（KD小于0.25 × 106），10-9 M）夹心ELISA更灵敏，检测0.1-1.0 μ g ml-1 IG，只要使用抗轻链MAb的缀合物。利用这些ELISA技术，分别对牛IgM、伊加、IgG 1和IgG 2特异性的4对单克隆抗体在来自100多头不同品种的牛的血清中进行筛选，以确定是否有检测到多态性表位。显示MAb IL-A30、IL-A60、IL-A66、IL-A71、IL-A72、IL-A73和IL-A74识别其各自重链上的单态决定簇。相比之下，在μ-通过MAb IL-A50对重链的扩增，发现其是等位的，并且在单个基因（可能是Cu）的控制下遗传。
A panel of 10 monoclonal antibodies specific for bovine immunoglobulins M, A, G1, G2 and light chains were produced and enzyme-linked immunosorbent assays developed to measure Ig levels in body fluids and culture supernatants using this panel of MAbs. An inhibition ELISA was accurate and sensitive for MAbs of high affinity, detecting levels as low as 10 ng ml-1 of IgM using a high-affinity MAb, IL-A50 (dissociation constant = 1.3 .times. 10-11 M). For MAbs of lower affinity (KD of less than 0.25 .times. 10-9 M) a sandwich ELISA was more sensitive, detecting 0.1-1.0 .mu.g ml-1 Ig, provided a conjugate of an anti-light chain MAb was used. Using these ELISA techniques, four pairs of MAbs specific for bovine IgM, IgA, IgG1 and IgG2 respectively, were screened on sera from over 100 cattle of different breeds to determine whether any detected a polymorphic epitope. MAbs IL-A30, IL-A60, IL-A66, IL-A71, IL-A72, IL-A73 and IL-A74 were shown to recognise monomorphic determinants on their respective heavy chains. In contrast, the epitope recognised on the .mu.-heavy chain by MAb IL-A50, which had previously been shown to be polymorphic, was found t o be allelic and inherited under the control of a single gene, probably Cu.