A novel rabbit immunospot array assay on a chip allows for the rapid generation of rabbit monoclonal antibodies with high affinity

一种新型的芯片兔免疫斑点阵列检测方法可以快速生成高亲和力的兔单克隆抗体

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作者:Tatsuhiko Ozawa, Xiuhong Piao, Eiji Kobayashi, Yue Zhou, Hiroaki Sakurai, Tsugunobu Andoh, Aishun Jin, Hiroyuki Kishi, Atsushi Muraguchi

Abstract

Antigen-specific rabbit monoclonal antibodies (RaMoAbs) are useful due to their high specificity and high affinity, and the establishment of a comprehensive and rapid RaMoAb generation system has been highly anticipated. Here, we present a novel system using immunospot array assay on a chip (ISAAC) technology in which we detect and retrieve antigen-specific antibody-secreting cells from the peripheral blood lymphocytes of antigen-immunized rabbits and produce antigen-specific RaMoAbs with 10(-12) M affinity within a time period of only 7 days. We have used this system to efficiently generate RaMoAbs that are specific to a phosphorylated signal-transducing molecule. Our system provides a new method for the comprehensive and rapid production of RaMoAbs, which may contribute to laboratory research and clinical applications.

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