Monoclonal antibody specific for alpha 2-->8-linked oligo deaminated neuraminic acid (KDN) sequences in glycoproteins. Preparation and characterization of a monoclonal antibody and its application in immunohistochemistry.
Two particular types of sialoglycoproteins have been detected in fish: polysialoglycoproteins containing alpha 2-->8-linked polysialic acid (-->8Neu5Gc alpha 2-->)n present in unfertilized Salmonidae fish eggs, and glycoproteins bearing oligo/polymers of deaminated neuraminic acids (KDN) found in the vitelline envelope of the eggs and ovarian fluid. We report the preparation and characterization of a monoclonal antibody specifically recognizing oligo/polymers of KDN sequences in glycoproteins and its application in immunohistochemistry. Fusion of spleen cells from a BALB/c mouse immunized with a KDN-rich glycoprotein (KDN-gp) containing (-->8KDN alpha 2-->)n-->6(KDN alpha 2-->3Gal beta 1-->3G alpha lNA-c alpha 1-->3) GalNAc alpha 1-->residues, with mouse myeloma cells yielded a hybrid cell line producing a monoclonal antibody that bound to KDN-gp, but not to KDN-gp depleted of KDN residues. The specificity of the monoclonal antibody, designated mAb.kdn8kdn, was determined by an enzyme-linked immunosorbent assay using KDN-gp samples that varied in KDN content. These antigens were prepared by the selective removal of KDN residues from the native KDN-gp. The mAb.kdn8kdn reacted most strongly with the intact KDN-gp and less strongly with KDN-gp samples containing decreased numbers of KDN residues.(ABSTRACT TRUNCATED AT 250 WORDS)
Kanamori
A
A
Department of Biophysics and Biochemistry, Graduate School of Science, University of Tokyo, Japan.
Inoue
S
S
Xulei
Z
Z
Zuber
C
C
Roth
J
J
Kitajima
K
K
Ye
J
J
Troy
F A
FA
Inoue
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002241:Carbohydrates / Q000032:analysis; D003641:Deamination; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006023:Glycoproteins / Q000276:immunology*; D006825:Hybridomas; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009438:Neuraminic Acids / Q000276:immunology*; D009841:Oligonucleotides / Q000276:immunology*; D017686:Oncorhynchus mykiss; D016612:Paraffin Embedding; D051381:Rats; D013154:Spleen / Q000166:cytology / Q000187:drug effects
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D006023:Glycoproteins; D009438:Neuraminic Acids; D009841:Oligonucleotides