Selective and Cleavable Extraction of Sialo-glycoproteins by Disulfide-Linked Amino-oxy-Functionalized Fe₃O₄ Magnetic Nanoparticles

The low abundance of sialo-glycoprotein hampered the separation, enrichment, and analysis of sialo-glycoproteins, which are critical for studying their functions. Here, we designed cleavable amino-oxy functionalized magnetic materials and employed to fast and selective isolate sialo-glycoproteins. This includes the ligation of disulfide-linked amino-oxy-functionalized magnetic nanoparticles with periodate-treated glycoproteins or cells, followed by magnetic separation. A reductive reagent could release the sialo-glycoproteins with small molecular fragments on the terminal of glycan chains, and the sialo-glycoproteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. On-bead digestion of the peptides were analyzed by tandem mass spectrometry. The results indicated that this method could selectively separate the majority of cell-surface sialo-glycoproteins

Selective and Cleavable Extraction of Sialo-glycoproteins by Disulfide-Linked Amino-oxy-Functionalized Fe₃O₄ Magnetic Nanoparticles

The low abundance of sialo-glycoprotein hampered the separation, enrichment, and analysis of sialo-glycoproteins, which are critical for studying their functions. Here, we designed cleavable amino-oxy functionalized magnetic materials and employed to fast and selective isolate sialo-glycoproteins. This includes the ligation of disulfide-linked amino-oxy-functionalized magnetic nanoparticles with periodate-treated glycoproteins or cells, followed by magnetic separation. A reductive reagent could release the sialo-glycoproteins with small molecular fragments on the terminal of glycan chains, and the sialo-glycoproteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis. On-bead digestion of the peptides were analyzed by tandem mass spectrometry. The results indicated that this method could selectively separate the majority of cell-surface sialo-glycoproteins

Additional file 1: of Variants of human papillomaviruses 16 (HPV16) in Uigur women in Xinjiang, China

Phylogenetic trees of HPV16 E6, E7 and L1 variants based on Neighbor Joining Molecular Phylogenetic analysis. Figure S1. Neighbor Joining Molecular Phylogenetic analysis using 43 nucleotide sequences of HPV16 E6 gene. Phylogenetic studies were performed on E6 nucleotide sequence alignment of 477 positions from each case, which was constructed by the neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package. Bootstrap proportions were calculated with 1000 replicates. Study sequences are labeled in KT GenBank accession numbers, others are reference GenBank sequences. E, European variant; Ep, European prototype; As, Asia lineage; AA, Asian American lineage; Af, African lineage. Figure S2. Neighbor Joining Molecular Phylogenetic analysis using 43 nucleotide sequences of HPV16 E7 gene. Phylogenetic studies were performed on E7 nucleotide sequence alignment of 297 positions from each case, which was constructed by the neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package. Bootstrap proportions were calculated with 1000 replicates. Study sequences are labeled in KT GenBank accession numbers, others are reference GenBank sequences. E, European variant; Ep, European prototype; As, Asia lineage; AA, Asian American lineage; Af, African lineage. Figure S3.  Neighbor Joining Molecular Phylogenetic analysis using 39 nucleotide sequences of HPV16 L1 gene. Phylogenetic studies were performed on partial L1 nucleotide sequence alignment of 369 positions from each case, which was constructed by the neighbor joining method and the Kimura 2-Parameter model by MEGA 6 package. Bootstrap proportions were calculated with 1000 replicates. Study sequences are labeled in KT GenBank accession numbers, others are reference GenBank sequences. E, European variant; Ep, European prototype; As, Asia lineage; AA, Asian American lineage; Af, African lineage. (PDF 136 kb