Detection and isolation of anti-hapten antibody-secreting cells by cellular affinity matrix technology

被引：1

作者：

Shimizu, Takeyuki ^{[1
]}

Azuma, Takachika ^{[2
]}

机构：

[1] Kochi Univ, Kochi Med Sch, Dept Immunol, Nankoku, Kochi 7838505, Japan

[2] Tokyo Univ Sci, RIBS, Lab Struct Immunol, Div Bioinformat, Noda, Chiba 2780022, Japan

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2015年 / 422卷

基金：

日本学术振兴会;

关键词：

Antibody; Plasma cells; Affinity matrix; Hapten; Flow cytometry; MEMORY B-CELLS; PLASMA-CELLS; CYTOKINE SECRETION; SECONDARY RESPONSE; NP ANTIBODIES; C57BL/6; MICE; ANTIGEN; MATURATION; GENERATION; ENRICHMENT;

D O I：

10.1016/j.jim.2015.04.006

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We developed a method to detect and isolate plasma cells that produce antigen-specific antibodies. An affinity matrix of hapten was constructed on a cell surface, and subsequent incubation allowed cells to secrete antibodies. Anti-hapten antibodies preferentially bound to the affinity matrix on the cells from which they were secreted. We showed that the combination of surface biotinylation and streptavidin which was conjugated with a high valence of hapten was suitable for sensitive detection of antibody binding. Using this protocol, anti-hapten plasma cells from immunized mouse spleen were detected and enriched by flow cytometry. This method allows for isolation of intact plasma cells according to the antibody specificity and may be useful for highly efficient and precise analysis of an antibody repertoire. (C) 2015 Elsevier B.V. All rights reserved.

引用

页码：80 / 86

页数：7

共 22 条

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