研究报告：单克隆抗体的鉴定及检测鸡白细胞介素 7 的夹心 ELISA 方法的建立
Research Note: Characterization of monoclonal antibodies and development of sandwich ELISA for detecting chicken IL7.
机构信息
Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, U.S. Department of Agriculture-ARS, Beltsville, MD 20705, USA.
Department of Animal Science and Technology, Chung-Ang University, Anseong-si, Gyeonggi-do 17546, Republic of Korea.
出版信息
Poult Sci. 2021 Mar;100(3):100940. doi: 10.1016/j.psj.2020.12.037. Epub 2021 Jan 5.
IL7 is a hematopoietic growth factor required for development and maintenance of lymphocytes including T cells, B cells, and natural killer cells. Recently, chicken IL7 (chIL7) has been cloned and studied in viral and parasite infection models. However, no monoclonal antibodies (mAb) that specifically detect chIL7 have been developed so far. In this study, recombinant chIL7 that expressed for immunization and mAb against chIL7 were developed and characterized to assess their immunologic properties. Five mAb exhibiting specific binding to chIL7 were generated and investigated for their applicability by Western blot, ELISA, and neutralization assays. A sandwich ELISA mAb pair that enables the measurement of chIL7 protein levels in biological samples from Eimeria-infected chickens was identified and several mAb neutralized chicken primary thymocyte proliferation mediated by chIL7. The mAb developed in this study will be valuable reagents for fundamental and applied immunological studies in poultry.
白细胞介素 7（IL7）是一种造血生长因子，对于淋巴细胞（包括 T 细胞、B 细胞和自然杀伤细胞）的发育和维持是必需的。最近，鸡白细胞介素 7（chIL7）已在病毒和寄生虫感染模型中被克隆和研究。然而，目前还没有开发出专门检测 chIL7 的单克隆抗体（mAb）。在这项研究中，我们开发并鉴定了用于免疫的重组 chIL7 和针对 chIL7 的 mAb，以评估其免疫特性。生成了 5 株特异性结合 chIL7 的 mAb，并通过 Western blot、ELISA 和中和测定来研究其适用性。鉴定出了一对用于夹心 ELISA 的 mAb 对，可用于检测感染艾美耳球虫的鸡生物样本中的 chIL7 蛋白水平，并且有几个 mAb 中和了 chIL7 介导的鸡原代胸腺细胞增殖。本研究中开发的 mAb 将是家禽基础和应用免疫学研究的有价值的试剂。
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