一系列针对血型糖蛋白A的新型单克隆抗体的精细表征。

Fine characterization of a series of new monoclonal antibodies directed against glycophorin A.

作者信息

Rasamoelisolo M, Czerwinski M, Bruneau V, Lisowska E, Blanchard D

机构信息

Etablissement de Transfusion Sanguine de Loire Atlantique/Vendée, Nantes, France.

出版信息

Vox Sang. 1997;72(3):185-91. doi: 10.1046/j.1423-0410.1997.7230185.x.

DOI:10.1046/j.1423-0410.1997.7230185.x

PMID:9145491

Abstract

OBJECTIVES

Glycophorins A (GPA) and B (GPB) are the major sialoglycoproteins of the human erythrocyte (RBC) membrane. To prepare tools for the analysis of GPA and GPB, we produced a series of new monoclonal antibodies (mAbs) that identified epitopes of GPA.

METHODS

Seven murine monoclonal antibodies directed to glycophorin A (GPA) were fully characterized by agglutination of untreated and enzyme-treated human erythrocytes, inhibition of agglutination using chemically modified glycophorins and peptides from GPA, immunoblotting, and binding to synthetic peptides on plastic pins.

RESULTS

The antibodies identify epitopes located on four different portions of GPA. (1) NaM13-6D2 binds to the N-terminal portion of GPA and GPB carrying the N blood group antigen; (2) NaM26-3F4 recognizes the homologous portion of GPA and GPB corresponding to their amino acids 6-26; (3) NaM10-2H12, NaM16-IB10 and NaM10-6G4 are specific for the amino acid sequence 38-45 of GPA; and (4) NaM37-5F4 and NaM13-4E4 bind to the amino acid residues 119-124 located on the intracellular ponion of GPA.

CONCLUSION

These antibodies represent precise tools to investigate GPA and related molecules in different cells and tissues.

摘要

目的

血型糖蛋白A（GPA）和血型糖蛋白B（GPB）是人类红细胞（RBC）膜的主要唾液酸糖蛋白。为制备用于分析GPA和GPB的工具，我们制备了一系列可识别GPA表位的新型单克隆抗体（mAb）。

方法

通过未处理和酶处理的人类红细胞凝集、使用化学修饰的血型糖蛋白和GPA肽抑制凝集、免疫印迹以及与塑料针上的合成肽结合，对7种针对血型糖蛋白A（GPA）的鼠单克隆抗体进行了全面表征。

结果

这些抗体识别位于GPA四个不同部分的表位。（1）NaM13 - 6D2与携带N血型抗原的GPA和GPB的N端部分结合；（2）NaM26 - 3F4识别GPA和GPB对应于其氨基酸6 - 26的同源部分；（3）NaM10 - 2H12、NaM16 - IB10和NaM10 - 6G4对GPA的氨基酸序列38 - 45具有特异性；（4）NaM37 - 5F4和NaM13 - 4E4与位于GPA胞内部分的氨基酸残基119 - 124结合。