A Novel Rabbit Immunospot Array Assay on a Chip Allows for the Rapid Generation of Rabbit Monoclonal Antibodies with High Affinity

Tatsuhiko Ozawa; Xiuhong Piao; Kobayashi, Eiji; Yue Zhou; Sakurai, Hiroaki; Andoh, Tsugunobu; Jin, Aishun; Kishi, Hiroyuki; Muraguchi, Atsushi
December 2012
PLoS ONE;Dec2012, Vol. 7 Issue 12, p1
Academic Journal
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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