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

被引：22

作者：

Ozawa, Tatsuhiko ^{[1
]}

Piao, Xiuhong ^{[1
]}

Kobayashi, Eiji ^{[1
]}

Zhou, Yue ^{[2
]}

Sakurai, Hiroaki ^{[2
]}

Andoh, Tsugunobu ^{[3
]}

Jin, Aishun ^{[1
,4
]}

Kishi, Hiroyuki ^{[1
]}

Muraguchi, Atsushi ^{[1
]}

机构：

[1] Toyama Univ, Grad Sch Med & Pharmaceut Sci, Dept Immunol, Toyama 930, Japan

[2] Toyama Univ, Grad Sch Med & Pharmaceut Sci, Dept Canc Cell Biol, Toyama 930, Japan

[3] Toyama Univ, Grad Sch Med & Pharmaceut Sci, Dept Appl Pharmacol, Toyama 930, Japan

[4] Harbin Med Univ, Coll Basic Med Sci, Dept Immunol, Harbin, Peoples R China

来源：

PLOS ONE | 2012年 / 7卷 / 12期

关键词：

SINGLE-CELL; B-CELLS; MOUSE HYBRIDOMAS; SECRETING CELLS; ANTIGEN; GROWTH; TAK1; TAB1; HETEROHYBRIDOMAS; PHOSPHORYLATION;

D O I：

10.1371/journal.pone.0052383

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页数：9

共 41 条

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