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- Title
Production and characterization of monoclonal antibodies specific to lactotriaosylceramide.
- Authors
Nozaki, Hirofumi; Yanagida, Mayumi; Koide, Ken-ichi; Shiotani, Kazusa; Kinoshita, Mikio; Kobayashi, Yoshiyasu; Watarai, Shinobu; Nakamura, Kazuo; Suzuki, Akemi; Ariga, Toshio; Kushi, Yasunori
- 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 (Lc3Cer). These obtained hybridoma cells, specific to Lc3Cer, 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 Lc3Cer 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.
- Subjects
MONOCLONAL antibodies; IMMUNOGLOBULIN G; IMMUNOGLOBULIN M; AMINO acids; ACUTE leukemia; CELL lines
- Publication
Glycobiology, 2010, Vol 20, Issue 12, p1631
- ISSN
0959-6658
- Publication type
Article
- DOI
10.1093/glycob/cwq117