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Production of a monoclonal antibody against chicken immunoglobulin G: A valuable molecule with research and diagnostic applications

机译:抗鸡免疫球蛋白G的单克隆抗体的生产:具有研究和诊断应用价值的有价值的分子

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Monoclonal antibodies (MAbs) are invaluable molecules which have several advantages over polyclonal immunoglobulins (Igs) including consistency and higher specificity and hence can be used in biological researches, diagnosis and treatment of diseases. The present study was conducted to produce monoclonal antibody against chicken IgG. The IgG molecules were purified from chicken serum and used as antigens to immunize several mice. Thereafter, a well-immunized mouse was chosen and used for fusion process. After production of hybridoma cells, several rounds of cloning were carried out and produced MAbs were examined by various immunological assays including enzyme-linked immunosorbent assay (ELISA) and western and dot blotting. Assessment of grown hybridomas indicated that only one clone (5B8) has produced desired MAb against chicken IgG. Meanwhile, using an indirect ELISA, it was shown that this MAb successfully recognizes chicken IgG molecules attached to influenza virus nucleoprotein. Evaluation of cross reactivity of MAb 5B8 with several avian serum samples revealed that this molecule specifically identifies chicken antibody molecules. However, it also recognized turkey antibodies with less affinity. In addition to research applications like isolation and purification of chicken and turkey IgG molecules, such a MAb can be applied to design and development of various immunoassays (e.g. ELISA) in these avian species. Key Words: Chicken, ELISA, Immunoglobulin G, Monoclonal antibody.
机译:单克隆抗体(MAbs)是无价的分子,与多克隆免疫球蛋白(Igs)相比具有多个优势,包括一致性和更高的特异性,因此可用于生物学研究,疾病的诊断和治疗。进行本研究以产生抗鸡IgG的单克隆抗体。从鸡血清中纯化出IgG分子,并用作抗原来免疫几只小鼠。此后,选择免疫良好的小鼠并用于融合过程。产生杂交瘤细胞后,进行几轮克隆,并通过各种免疫学测定法检查产生的单克隆抗体,包括酶联免疫吸附测定法(ELISA)以及蛋白质印迹和斑点印迹法。对生长的杂交瘤的评估表明,只有一个克隆(5B8)产生了抗鸡IgG的所需单克隆抗体。同时,使用间接ELISA显示,该MAb成功地识别了附着于流感病毒核蛋白的鸡IgG分子。 MAb 5B8与几种禽类血清样品的交叉反应性评估显示,该分子可特异性鉴定鸡抗体分子。但是,它也以较低的亲和力识别火鸡抗体。除了诸如鸡和火鸡IgG分子的分离和纯化之类的研究应用外,此类MAb还可用于设计和开发这些禽类中的各种免疫测定法(例如ELISA)。关键词:鸡,ELISA,免疫球蛋白G,单克隆抗体。

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