Fluorescent glycans

Inject 1 p.L of the sample for PGC-HPLC analysis. Use both fluorescence and UV detection. Use the profiles as guidance for the following purification. Inject all of the sample into the same PGC-HPLC column using UV detection only. Collect the major peak as the fluorescent glycan-AEAB conjugate. 

Song X, Xia B, Lasanajak Y, Smith DF, Cummings RD (2008) Quantifiable fluorescent glycan microarrays. Glycoconj J 25 15-25... 

Song X, et al (2009) Novel fluorescent glycan microarray strategy reveals ligands for galectins. Chem Biol 16 36-47... 

Sawa M, Hsu TL, Itoh T et al (2006) Glycoproteomic probes for fluorescent imaging of fucosylated glycans in vivo. Proc Natl Acad Sci USA 103 12371-12376... 

Figure 1.116 Released glycans can be labeled with small fluorescent compounds containing amines for subsequent detection upon chromatographic separation. In the presence of sodium cyanoborohydride these compounds react at the reducing end of glycans to form secondary amine derivatives with characteristic spectral properties.

The labeled glycans may be analyzed by cell sorting after staining with fluorescently modified streptavidin. Alternatively, the cells may be lysed and the labeled glycans isolated using immobilized streptavidin. 

Bigge, J.C., Patel, T.P., Bruce, J.A., Goulding, P.N., Charles, S.M., and Parekh, R.B. (1995) Nonselective and efficient fluorescent labeling of glycans using 2-amino benzamide and anthranilic acid. Anal. Biochem. 230, 229-238. 

Fig. 1 Glycan array analysis of codakine as measured by fluorescence intensity (glycan array v3.0 at Consortium for Functional Glycomics). Purified codakine from white clams was purified with Alexa Fluor 488 Protein Labeling Kit (Molecular Probes , Invitrogen) and tested on glycan array v3.0 at Consortium for Functional Glycomics.

Figures Selective protein modification using a keto amino acid, p-acetyl-L-phenylalanine. (a) Labeling of fluorescein hydrazide to the Z domain protein. Only the mutant protein containing p-acetyl-L-phenylalanine was labeled and became fluorescent, (b) A general method for preparing glycoprotein mimetics with defined glycan structure.

Zhuang et al. [152] derivatized glycan samples with 8-aminopyrene-l,3,6-trisulfonic acid to get a charge for electrophoresis and a fluorescent label for detection. They analyzed glycan derivatives in the blood of cancer patients... 

Recent detection methods for glycan array include fluorescent assay, SPR, MALDI-TOF mass spectrometry, and nanoparticle assay. Fluorescence-based measurement is the prevalent principle for detecting binding to glycan microarrays. Rhodamine [9],... 

Another metal present in biological media in relatively high concentrations is iron. Most iron ions are strongly coordinated within proteins, although a small fraction of iron ions is loosely bound with carboxylates and phosphates, phospholipids, glycans, and sulphonates. This fraction is usually called labile iron and constitutes a pool of iron ions accessible for incorporation into functional metalloproteins. In contrast to closed shell cations, rather than increasing the fluorescence quantum... 

---