禽白血病毒单克隆抗体的特性

程由铨; Lucy F. lee; Robert F. Silva; Fugene J. Smith; Lyman B. Crittenden

禽白血病毒单克隆抗体的特性

1.
福建省农业科学院畜牧兽医研究所;
2.
美国农业部农业科学研究署地区家禽研究所

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出版历程
- 刊出日期: 1986-09-15

CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO AVIAN LEUKOSIS VIRUSES

1.
Institute of Animal Husbandry and Veterinary Medicine, Fujian Academy of Agricultural Sciences;
2.
U. S. Department of Agriculture, Agricultural Research Service, Regional Poultry Research Laboratory

摘要

摘要: 建立了分泌抗禽白血病毒(ALV)结构蛋白P27和P19的单克隆抗体(McAb)杂交瘤细胞。酶联免疫吸附试验(ELISA),McAb-6AL20(属于IgG1)分别与外源性ALV:A.B.D亚群的RPL-40、AMV、RAV-2和Carr-Zilber RSV(CZ-RSV)发生特异性反应,而不与内源性ALV:RAV-O(E亚群)发生反应。McAb-6AL22(属于IgG1)除了上述特性外,还能与劳斯氏肉瘤病毒Prague株(Prc-RSV、属于C亚群)发生反应。这二个McAb均能与〔35S〕甲硫氨酸标记的RPL-40和RAV-1病毒的P19蛋白出现免疫沉淀。但是,它们不与RAV-O病毒的P19蛋白发生免疫沉淀。所以,McAb可用于区别ALV结构蛋白P19(亚群特异性位点的多肽)。应用这二个McAb进行ELISA和免疫沉淀与聚丙烯酰胺凝胶电泳试验可区分RPL-40和RAV-O感染。McAb-6AL42(属于IgG2a)可与以上几个亚群(A.B.C和D)的毒株发生反应,其ELISA的抗体滴度比与RAV-O(E亚群)毒株高1000倍以上。McAb-6AL42可与P27和pr76(群特异性抗原的前体蛋白)发生免疫沉淀。试验用McAb-6AL42作为第一抗体和兔抗P27血清的过氧化物酶标记物作为第二抗体(指示抗体)建立了双抗体夹心ELISA试验。用这种ELISA试验可区分未经稀释的卵蛋白中RAV-O和RPL-40群特异性抗原。

Abstract: Hybridoma cell lines secreting monoclonal antibody (MCA) to avian leukosis virus (ALV) structural proteins p27 and p19 have been established. MCA 6AL20 (IgG1 isotype) reacted with RPL-40, AMV, RAV-2 and Carr-Zilber RSV (CZ-RSV).representing exogenous ALV subgroups A, B and D, respectively, but not the endogenous virus RAV-0 (subgroup E) in an indirect enzyme-linked immunosorbent assay (ELISA). MCA 6AL22 reacted as above and, in addition, reacted with the Prague strain of Rous sarcoma virus (PrC-RSV), subgroup C. Both MCAs im-munoprecipitated p19 from 35S-methionine-labeled RPL-40 or RAV-1,but not RAV-O infected chi-cken embryo fibroblasts (CEF). They can be used to differentiate exogenous from endogenous (RAV-O) infection either in an indirect antibody ELISA or by immunoprecipitation. A third MCA, 6AL42 (IgG2a isotype), reacted with exogenous viruses at an antibody titer up to 1,000-fold higher than with endogenous subgroup E, RAV-O virus in indirect ELISAs. MCA 6AL42 immunoprecipitated p27 and the group-specific antigen precursor protein, pr76, from cells infected with RPL-40, RAV-1 or RAV-O. A double antibody sandwich ELISA, using 6AL42 as the primary binding antibody and conjugated rabbit anti-p27 as the reporter antibody, differentiated between endogenous RAV-O and exogenous p27 antigen in undiluted albumen samples.

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参考文献(8)

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