Conjugates-Effect-Ab-Affinity-v

In contrast, conjugation of less than five chelatesAb fragment randomly at lysine residues resulted in a three- to fivefold reduction in affinity. By a single Arg to Asn mutation, an N-linked glycosylation site similar to that of LL-2 was introduced in the FR-1 segment of a nonglycosylated, humanized anti-carcinoembryonic Ag (CEA) Ab, MN-14 (hMN-14). Glycosylation at the engineered carbohydrate-addition site was demonstrated by SDS-PAGE analysis. Neither glycosylation nor site-specific conjugation of chelate at the V kappa-appended carbohydrate moiety resulted in the loss of immunoreactivity. The glycosylated hMN-14 conjugate labeled efficiently with Characterization of the N-linked oligosaccharides from human chorionic gonadotropin expressed in the methylotrophic yeast Pichia pastoris.Human chorionic gonadotropin (hCG) is a heterodimeric, placental glycoprotein hormone involved in the maintenance of the corpus luteum during the first trimester of pregnancy.

Biologically active hCG has been successfully expressed in the yeast Pichia pastoris (phCG). In the context of structural studies and therapeutic applications of phCG, detailed information about its glycosylation pattern is a prerequisite. To this end N-glycans were released with peptide-N(4)-(N-acetyl-beta-glucosaminyl)asparagine amidase F and fractionated via anion-exchange chromatography (Resource Q) yielding both neutral (%) and charged, phosphate-containing (%) high-mannose-type structures. Subfractionations were carried out via normal phase (Lichrosorb-NH(2)) and high-pH anion-exchange (CarboPac PA-1) chromatography. Structural analyses of the released N-glycans were carried out by using HPLC profiling of fluorescent 2-aminobenzamide derivatives, MALDI-TOF mass spectrometry, and 0-MHz(1)H-NMR spectroscopy. Detailed neutral oligosaccharide structures, in the range of Man(8)GlcNAc(2) to Man(11)GlcNAc(2) including molecular isomers, could be established, and structures up to Man(15)GlcNAc(2) were indicated. Phosphate-containing oligosaccharides ranged from Man(9)PGlcNAc(2) to Man(13)PGlcNAc(2).

Human Milk Glycans -glycans were not detected. Profiling studies carried out on different production batches showed that the oligosaccharide structures are similar, but their relative amounts varied with the culturing Selective cleavage of glycosidic linkages studies with the polysaccharide component of Shigella dysenteriae type 6 lipopolysaccharide.The polysaccharide component obtained from the lipopolysaccharide of Shigella dysenteriae type 6 was subjected to milk hydrolysis with acid, and the products were fractionated on Sephadex G-. An acidic hexosaminoglycan and a core oligosaccharide fraction were obtained, the former containing D-glucose, D-galactose, 2-acetamido-2-deoxy-D-galactose (in the ratios 111), and an unidentified acidic component (X). 2'-Fucose lactose was N-deacetylated and then hydrolysed and deaminated to give 3-O-(2-amino-2-deoxy-beta-D-galactopyranosyl)-D-galactose (1), identified as the 2,5-anhydro-3-O-(6-O-alpha-D-galactopyranosyl-alpha-D-glucopyranosyl)talitol the polysaccharide and 3, together with that for the determination of linkage configurations by chromic anhydride oxidation, the hexosaminoglycan is considered to have the repeating structure (see article)516. Zentralbl Bakteriol Mikrobiol Hyg A Med Mikrobiol Infekt Parasitol. The lipopolysaccharide of escherichia coli C- studies on the anomeric configurations of the hexoses in the R1 core.

Lipopolysaccharide from E, coli C as well as lipopolysaccharides from submutants of E. coli with incomplete core structures in their lipopolysaccharides were isolated and quantitatively analyzed. Core oligosaccharides were isolated from lipopolysaccharides by acetic acid degradation and were purified by gel chromatography. The difference in molecular rotations of the core oligosaccharides from E. coli C and 6 submutants thereof with incomplete core structure were correlated to the differences in sugar compositions. The anomeric configurations have been deducted from the high or low contribution of each individual sugar to the molecular rotation of the core oligosaccharide from E. coli C.

The primary structure of the hexose region of the lipopolysaccharide from E. coli C is primary structure of the hexose region of the lipopolysaccharide from E. coli C is, see formula in text. The anomeric configurations of glucoses I, II, and III were confirmed by precipitation reactions of alkali treated lipopolysaccharides from E. coli C, C23. 1, and C21 with Concanavalin A.