Use of Butyl Acetate in Determination of Sialic Acid.

Miettinen, T. A.; Takki-Luukkainen, I.-T.; Smith-Kielland, I.; Sömme, R.; Stenhagen, Einar; Palmstierna, H.

doi:10.3891/acta.chem.scand.13-0856

Cited by 640 publications

(137 citation statements)

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“…Glycopeptides were desalted on a P-2 (Bio-Gel P-2/200-400 mesh, Bio-Rad, Richmond, CA, USA) gel filtration column (75 cm × 2.5 cm, total bed volume 368 ml), with 10 ml of Bio-Gel P-6 DG (Bio-Rad, Richmond, CA, USA) on the bottom to prevent clogging of the lower frit. The column was eluted with 50 mM ammonium bicarbonate, pH 7.0 at the flow rate of 0.5 ml/min and the fractions (12 ml) collected were analyzed colorimetrically for sialic acid (Miettinen and Takki-Luukkainen, 1959). The fractions containing sialylated glycopeptides were pooled, lyophilized with sodium bicarbonate (see earlier) and stored frozen.…”

Section: Preparation Of Poly-and Oligosaccharidesmentioning

confidence: 99%

High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines

Häyrinen¹,

Haseley²,

Talaga³

et al. 2002

Molecular Immunology

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Long-chain polysialic acid (PSA) is expressed on the vertebrate neural cell adhesion molecule (NCAM) during neuronal plasticity. Its structural similarity to the capsular PSAs of some pathogenic bacteria has hampered the development of polysaccharide vaccines against meningitis. The antibodies formed during immunization require a long epitope for binding, and cross-react with host tissue PSA. The nature of the epitope and possible external effectors involved are still unclear. We have evaluated the interaction of PSA with its antibody mAb735 by surface plasmon resonance. The influences of PSA chain length, pH, temperature, ionic environment, and polyamines were also determined.The antibody binding affinity was found to dramatically increase with PSA chain length. A sub-nanomolar dissociation constant (K D = 8.5 × 10 −10 M) was obtained for the binding of very long chain native MenB polysaccharides (∼200 Neu5Ac-residues). Colominic acid from Escherichia coli K1 (∼100 residues) and shorter polymers exhibited progressively weaker affinities. The antibody also bound tightly (K D ∼ 5 × 10 −9 M) to polysialylated glycopeptides from human embryonal brain. The effects of pH and ionic strength suggested that the interaction is largely electrostatic. Ca 2+ and Mn 2+ ions promoted the observed surface plasmon resonance response in a concentration dependent fashion. Spermine increased the response in a similar way. Our results suggest that divalent cations and polyamines may play significant role in the regulation of the PSA epitope presentation in vivo.

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Section: Preparation Of Poly-and Oligosaccharidesmentioning

confidence: 99%

High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines

Häyrinen¹,

Haseley²,

Talaga³

et al. 2002

Molecular Immunology

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“…The amount of gangliosides was determined as lipid-bound sialic acid using resorcinol/HCl reagent (Svennerholm, 1957) as modified by Miettinen & Takki-Lukkainen (1959).…”

Section: Methodsmentioning

confidence: 99%

Adhesion of K99 fimbriated Escherichia coli to pig intestinal epithelium: correlation of adhesive and non-adhesive phenotypes with the sialoglycolipid content

Seignole¹,

Mouricout²,

Duval-Iflah

et al. 1991

Journal of General Microbiology

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Evidence for the existence of two phenotypes of piglets born to experimental herds was obtained based on the susceptibility of intestinal brush borders to adhesion of K99-positive Escherichia cofi. The enterocytes of the K99-receptive piglets displayed a characteristic sialoglycolipid pattern, with a higher content of the monosialoglycolipids I13NeuGc-LacCer (GM~Gc), IV3NeuGc-nLcOse4Cer (SPGGc) and IV3NeuAc-nLcOse4Cer (SPG) and the oligosialogangliosides IV3NeuAc,I13NeuAc-GgOse,Cer (GDla), I13(NeuAc),-GgOse,Cer (GD2), I13(NeuAc)z-GgOse4Cer (GD1 b) and IV3NeuAc,I13(NeuAc)2-GgOse4Cer (GTlb) when compared to the gangliosides of non-receptive piglets. The gangliosides from enterocytes of the non-receptive piglets were mainly the monosialogangliosides 113 NeuAc-GgOse,Cer (GM2) and I13NeuAc-LacCer (GM3), only traces of the other sialoglycolipids being detected. Adhesion of 4C-labelled K99-positive E. cofi cells to the piglet small intestinal sialoglycolipids, as tested by the thin-layer chromatogram overlay assay, revealed that the receptive enterocyte membrane was richer in glycolipids containing K99 receptor structures than the non-receptive enterocyte.Adhesion of K99-positive E. cofi correlated with the degree of sialylation of the brush border glycolipids.

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“…Analytical methods 2.4. Methylation studies Total neuraminic acid was determined by the resorcinol method of Svennerholm [8], as modified by Miettinen and Takki-Luukkainen [9]. Free neuraminic acid was assayed by the thiobarbituric acid reaction of Warren [IO] .…”

Section: Methodsmentioning

confidence: 99%

Action of Clostridium perfringens neuraminidase on gangliosides GM₁ and GM₂ above and below the critical micelle concentration of substrate

Rauvala¹

1976

FEBS Letters

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Use of Butyl Acetate in Determination of Sialic Acid.

Cited by 640 publications

References 0 publications

High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines

High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines

Adhesion of K99 fimbriated Escherichia coli to pig intestinal epithelium: correlation of adhesive and non-adhesive phenotypes with the sialoglycolipid content

Action of Clostridium perfringens neuraminidase on gangliosides GM₁ and GM₂ above and below the critical micelle concentration of substrate

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Use of Butyl Acetate in Determination of Sialic Acid.

Cited by 640 publications

References 0 publications

High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines

High affinity binding of long-chain polysialic acid to antibody, and modulation by divalent cations and polyamines

Adhesion of K99 fimbriated Escherichia coli to pig intestinal epithelium: correlation of adhesive and non-adhesive phenotypes with the sialoglycolipid content

Action of Clostridium perfringens neuraminidase on gangliosides GM1 and GM2 above and below the critical micelle concentration of substrate

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Action of Clostridium perfringens neuraminidase on gangliosides GM₁ and GM₂ above and below the critical micelle concentration of substrate