title: A novel rabbit immunospot array assay on a chip allows for the rapid generation of rabbit monoclonal antibodies with high affinity.
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.
authors.affiliation: Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Ozawa
authors.forename: Xiuhong
authors.initials: X
authors.lastname: Piao
authors.forename: Eiji
authors.initials: E
authors.lastname: Kobayashi
authors.forename: Yue
authors.initials: Y
authors.lastname: Zhou
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Sakurai
authors.forename: Tsugunobu
authors.initials: T
authors.lastname: Andoh
authors.forename: Aishun
authors.initials: A
authors.lastname: Jin
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Muraguchi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D046228:Microarray Analysis / Q000379:methods*; D010455:Peptides / Q000276:immunology / Q000378:metabolism; D010766:Phosphorylation; D011817:Rabbits; D015398:Signal Transduction; D013997:Time Factors
chemical_list: D000911:Antibodies, Monoclonal; D010455:Peptides