RRC ID 38258
Author Nozaki H, Yanagida M, Koide K, Shiotani K, Kinoshita M, Kobayashi Y, Watarai S, Nakamura K, Suzuki A, Ariga T, Kushi Y.
Title Production and characterization of monoclonal antibodies specific to lactotriaosylceramide.
Journal Glycobiology
Abstract We have established hybridoma cell lines producing monoclonal antibodies (mAbs) directed to N-acetylglucosaminylβ1-3galactose (GlcNAcβ1-3Gal) residue by immunizing BALB/c mice with lactotriaosylceramide (Lc(3)Cer). These obtained hybridoma cells, specific to Lc(3)Cer, were dual immunoglobulin (Ig)-producing cells which secreted both IgM and IgG molecules as antibodies. The established mAbs are able to react with not only Lc(3)Cer but also GlcNAcβ1-3-terminal glycosphingolipids (GSLs) despite branching or lactosamine chain lengths and human transferrin with terminal GlcNAc residues. Comparison of the variable regions of the cloned IgM and IgG by reversed transcription-polymerase chain reaction analysis confirmed that the variable regions determine the specificity, the other amino acids are conserved, and these mAbs are encoded by J558 and Vκ-21family genes. Furthermore, we have analyzed the expression of GSLs with GlcNAcβ1-3 epitope in acute leukemia cell lines and mouse fetal tissues using these mAbs, in which antigens were distributed comparatively. These mAbs are useful for studying the precise distribution of GlcNAcβ1-3Gal-terminating GSL expression in tissues as well as for detecting GSLs carrying terminal GlcNAcβ1-3Gal carbohydrate structure.
Volume 20(12)
Pages 1631-42
Published 2010-12
DOI 10.1093/glycob/cwq117
PII cwq117
PMID 20693232
MeSH Animals Antibodies, Monoclonal, Murine-Derived / biosynthesis Antibodies, Monoclonal, Murine-Derived / genetics Antibodies, Monoclonal, Murine-Derived / immunology* Antibody Specificity / genetics Antibody Specificity / immunology* Female HL-60 Cells Humans Immunoglobulin G / biosynthesis Immunoglobulin G / genetics Immunoglobulin G / immunology* Immunoglobulin M / biosynthesis Immunoglobulin M / genetics Immunoglobulin M / immunology* Immunoglobulin Variable Region / biosynthesis Immunoglobulin Variable Region / genetics Immunoglobulin Variable Region / immunology* K562 Cells Lactosylceramides / biosynthesis Lactosylceramides / genetics Lactosylceramides / immunology* Mice Mice, Inbred BALB C U937 Cells
IF 3.664
Times Cited 6
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells