Monoclonal antibodies directed to the synthetic carbohydrate antigen Ley.
Tetrasaccharide Fuc alpha 1-2Gal beta 1-4(Fuc alpha 1-3)GlcNAc is known as carbohydrate determinant of cancer- and AIDS-associated antigen Lewisy (Ley). Synthetic antigen to generate mouse monoclonal antibodies (MAbs) directed to Ley was prepared and constructed as a spacer-armed tetrasaccharide coupled with lipophilized polymer, Ley-PAA-PE, where PAA is a 30-kD polyacrylamide and PE is phosphatidylethanolamine. An efficient immune response was provided by using Ley-PAA-PE adsorbed on Salmonella minnesota. Positive hybridomas were screened by enzyme-linked immunosorbent assay (ELISA) using Ley-PAA as a coating agent. An inhibitory version of the same test system showed absolute specificity of two MAbs: only hapten Ley and Ley-PAA were strong inhibitors, in contrast to Leb, tri- and disaccharidic fragments of the mentioned tetrasaccharides, as well as their PAA-conjugates. MAbs obtained against synthetic antigen specifically stained the Ley (+) cell line A431.
Vlasova
E V
EV
Shemyakin Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
Byramova
N E
NE
Tuzikov
A B
AB
Zhigis
L S
LS
Rapoport
E M
EM
Khaidukov
S V
SV
Bovin
N V
NV
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000276:immunology*; D002240:Carbohydrate Sequence; D005260:Female; D005434:Flow Cytometry; D016256:Lewis X Antigen / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D015295:Antigens, Tumor-Associated, Carbohydrate; D016256:Lewis X Antigen