Lectin affinity electrophoresis for the separation of fluorescently labeled sugar derivatives.
Lectin affinity electrophoresis was applied to the separation of charged, fluorescent conjugates of disaccharides. Four fluorescent conjugates were prepared by reductive amination of alpha-D-Man-(1----3)-D-Man, alpha-D-Gal-(1----4)-D-Glc, alpha-D-Gal-(1----6)-D-Glc, and beta-D-Gal-(1----4)-D-Glc in the presence of 7-amino-1,3-naphthalenedisulfonic acid. These charged fluorescent-disaccharide conjugates all have identical molecular weight and in the absence of conconavalin A lectin failed to separate either by agarose or by polyacrylamide gel electrophoresis. In the presence of either free or immobilized concanavalin A, agarose gel electrophoresis and polyacrylamide gel electrophoresis could separate the fluorescent conjugate of alpha-D-Man-(1----3)-D-Man from that of alpha-D-Gal-(1----4)-D-Gal, alpha-D-Gal-(1----6)-D-Glc, and beta-D-Gal-(1----4)-D-Glc.
Lee
K B
KB
Division of Medicinal and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242.
Kim
Y S
YS
Linhardt
R J
RJ
D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000737:chemistry; D002240:Carbohydrate Sequence; D004187:Disaccharides / Q000737:chemistry / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D006001:Glycoconjugates / Q000737:chemistry / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D012685:Sepharose / Q000031:analogs & derivatives
D004187:Disaccharides; D005456:Fluorescent Dyes; D006001:Glycoconjugates; C028326:concanavalin A-sepharose; C033153:amido-G-acid; D012685:Sepharose; D015081:2-Naphthylamine