Transfer of 131I and fluoresceinyl sialic acid derivatives into the oligosaccharide chains of IgG: a new method for site-specific labeling of antibodies.
Biochemical modifications of IgG can help to avoid damages caused by oxidation or reduction. Terminal groups of the saccharide structures, located in the Fc-portion of IgG molecules, were modified by enzymatic reactions. IgG was pretreated with sialidase, to cleave bound sialic acid, and with galactosyltransferase, to increase the number of acceptor sites for transfer reactions. Afterward, modified sialic acid derivatives were transferred enzymatically into the oligosaccharide chains of IgG. Labeling was possible with sialic acids modified in either position 5 or position 9. The usefulness of this method was demonstrated for radioactive and fluoresceinylated reagents, with yields up to 90% in 1 h. Immunological investigations have shown no influence on the immunoreactivity by the described modification of saccharide structures.
Schwarz
U
U
Institut für Analytische Chemie, Technische Universität Dresden, Germany.
Wunderlich
G
G
Brossmer
R
R
D000818:Animals; D019793:Fluorescein; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology; D007457:Iodine Radioisotopes; D007553:Isotope Labeling / Q000379:methods*; D051379:Mice; D009844:Oligosaccharides / Q000737:chemistry*; D012794:Sialic Acids / Q000737:chemistry*
D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D009844:Oligosaccharides; D012794:Sialic Acids; D019793:Fluorescein