MONOCLONAL-ANTIBODIES SPECIFIC FOR ADENOVIRUS EARLY REGION 1A PROTEINS - EXTENSIVE HETEROGENEITY IN EARLY REGION 1A PRODUCTS
MONOCLONAL-ANTIBODIES SPECIFIC FOR ADENOVIRUS EARLY REGION 1A PROTEINS - EXTENSIVE HETEROGENEITY IN EARLY REGION 1A PRODUCTS
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DOI：
10.1128/jvi.55.3.533-546.1985
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发表时间：
1985-01-01
影响因子：
5.4
通讯作者：
SCHLEY, C
中科院分区：
文献类型：
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作者：
HARLOW, E;FRANZA, BR;SCHLEY, C
用细菌trpE-E1 A融合蛋白免疫BALB/c小鼠，制备分泌腺病毒E1 A蛋白特异性单克隆抗体的杂交瘤。该蛋白由一个杂交基因编码，该基因连接了大肠杆菌trpE基因的一部分和E1 A 13 S mRNA的互补DNA拷贝。分泌抗体5，129的杂交瘤（83）对融合蛋白的E1 A部分具有特异性。只有12个单克隆抗体可以有效地免疫沉淀E1 A多肽从受感染细胞的去污剂裂解物。在一维十二烷基硫酸钠-聚丙烯酰胺凝胶和二维等电聚焦聚丙烯酰胺凝胶上分析E1 A多肽。被单克隆抗体特异性免疫沉淀的E1 A蛋白在大小和电荷上是异质的，并且可以被解析成约60种多肽。这种异质性不仅是由于多种E1 A mRNA的合成，而且至少部分是由于翻译后修饰。几种单克隆抗体根据抗体与抗原结合的能力将E1 A多肽分为免疫学亚类。在这些单克隆抗体中，有2种与由13 S E1 A mRNA合成的多肽结合，但不与其它E1 A蛋白结合。
Hybridomas secreting monoclonal antibodies specific for the adenovirus early region 1A (E1A) proteins were prepared from BALB/c mice immunized with a bacterial trpE-E1A fusion protein. This protein is encoded by a hybrid gene that joins a protion of the Esherichia coli trpE gene and a complementary DNA copy of the E1A 13S mRNA. Hybridomas (83) that secrete antibodies 5,129 are specific for the E1A portion of the fusion protein. Only 12 of the monoclonal antibodies can efficiently immunoprecipitate E1A polypeptides from detergent lysates of infected cells. E1A polypeptides were analyzed on 1-dimensional, sodium dodecyl sulfate-polyacrylamide gels and 2-dimensional isoelectric focusing polyacrylamide gels. The E1A proteins that are specifically immunoprecipitated by the monoclonal antibodies are heterogeneous in size and charge and can be resolved into .apprx. 60 polypeptide species. This heterogeneity is due not only to synthesis from multiple E1A mRNA species, but also at least in part to post-translational modification. Several of the monoclonal antibodies divide the E1A polypeptides into immunological subclases based on the ability of the antibodies to bind to the antigen. Of these monoclonal antibodies, 2 bind to the polypeptides synthesized from the 13S E1A mRNA, but not to other E1A proteins.