抗原抗体共免疫制备乙肝e抗原特定表位的抗体以及其在检测中的应用

发布时间：2019-04-21 11:24

【摘要】：近年来,肝病已越来越成为当今威胁人类健康的主要疾病之一,病毒性肝炎中尤以乙型肝炎对人类影响最大,乙型肝炎病毒(HBV)感染是一个严重的全球性公共健康问题,成为了当前的一种世界性疾病。由于乙肝影响范围的广泛性,危害的严重性,乙型肝炎诊断试剂作为专用于乙肝患者各种生理指标的检测、乙肝的诊断和治疗过程监测的生物学和化学试剂,具有广泛的应用范围和社会需求。本研究就是在以前研究的基础上,开发出针对乙肝e抗原的抗体。E抗原全长159个氨基酸,对它的抗原决定簇的性质的研究也已经有30多年的历史,目前比较公认的是它有三个主要抗原决定簇,两个线性的,一个构型的,其中一个主要线性抗原决定簇是HBeβ,定位在120-133氨基酸上,这是一个优势抗原决定簇。由于这个优势决定簇的存在,传统的免疫方法只能获得针对它的抗体。本研究采用改变常规的免疫方法而采用封闭优势决定簇的策略期望获得针对非主要抗原决定簇的抗体,封闭的方法是用目前已经获得的针对这个主要决定簇的抗体和抗原共免疫。通过免疫方法的改变,最后得到了两个单克隆抗体-S-29-3和S-72-3。对它们的结合表位研究表明,S-29-3识别的表位是一个包含HBeβ这段序列在内的构型表位;S-72-3识别的是一个包含了一个很长序列氨基酸(2-118)的构型表位。Biosensor亲和力检测显示他们都具有很高的亲和常数,用它们代替进口产品进行配对检测重组抗原,具有和商品化的配对抗体lpa相似的灵敏度。
[Abstract]:In recent years, liver disease has become one of the main diseases threatening human health more and more, especially hepatitis B has the greatest impact on human beings. Hepatitis B virus (HBV) infection is a serious global public health problem. It has become a worldwide disease. Because of the extensive scope of hepatitis B infection and the severity of the harm, hepatitis B diagnostic reagent is used as a biological and chemical reagent for the detection of various physiological indexes of hepatitis B patients, the diagnosis and treatment of hepatitis B, With a wide range of applications and social needs. On the basis of previous studies, we have developed antibodies against hepatitis B e antigen. The whole length of E antigen is 159 amino acids, and the properties of its antigenic determinants have been studied for more than 30 years. It is generally recognized that it has three main antigenic determinants, two linear and one configuration, one of which is HBe 尾, which is located on 120x133 amino acids, which is a dominant antigenic determinant. Because of the existence of this dominant determinant, traditional immune methods can only obtain antibodies against it. In this study, we used the strategy of blocking dominant determinants to obtain antibodies against non-major antigenic determinants by changing the conventional immune methods and using the strategy of blocking dominant determinants. The blocking method is to co-immunize with antibodies and antigens that are currently available against this major determinant. Through the change of immune method, two monoclonal antibodies-Sx293 and SJ72m3 were obtained. The results were as follows: (1). The study of their binding epitopes showed that the epitope recognized by Sx293 was a configuration epitope containing the sequence of HBe 尾. The biosensor affinity test shows that they all have a high affinity constant, so they are used to pair the recombinant antigen instead of the imported product, and the biosensor affinity test shows that they all have high affinity constant, and that the recombinant antigen contains a long sequence of amino acids (2 渭 118), and the biosensor affinity test shows that they all have very high affinity constants. It has a sensitivity similar to that of commercial pairing antibody lpa.
【学位授予单位】：中国科学院研究生院（上海生命科学研究院）
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

【共引文献】