sialidase  (New England Biolabs)


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    Name:
    alpha 2 3 6 8 9 Neuraminidase A
    Description:
    alpha 2 3 6 8 9 Neuraminidase A 4 000 units
    Catalog Number:
    P0722L
    Price:
    278
    Category:
    Glycosidases
    Size:
    4 000 units
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    Structured Review

    New England Biolabs sialidase
    alpha 2 3 6 8 9 Neuraminidase A
    alpha 2 3 6 8 9 Neuraminidase A 4 000 units
    https://www.bioz.com/result/sialidase/product/New England Biolabs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sialidase - by Bioz Stars, 2021-09
    94/100 stars

    Images

    1) Product Images from "Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration"

    Article Title: Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration

    Journal: eLife

    doi: 10.7554/eLife.61552

    Principal findings and conceptual model. ( A ) ACE2-Fc binding was measured to wild-type or glycoEnzyme-KO 293 T cells expressing Spike. Sialidase treatment of cells was performed in some cases. Similar studies also measured S1-Fc and RBD-Fc binding to cell-surface expressed ACE2. ( B ) SARS-CoV-2 pseudovirus (bearing Spike-WT, Spike-mut, Spike-delta variants) were generated in wild-type or glycoEnzyme-KO 293Ts, in the presence and absence of kifunensine. Main results of binding ( A ) and viral entry ( B ) assay are listed. ( C ) Conceptual model shows that kifunensine can induce S1-S2 site proteolysis on Spike-WT and Spike-mut virus, but not Spike-delta virus. This proteolysis reduces RBD presentation and attenuates viral entry into 293T/ACE2. Without affecting S1-S2 cleavage, kifunensine also partially reduced Spike-delta pseudovirus entry function. The data suggest additional roles for Spike N-glycans during viral entry.
    Figure Legend Snippet: Principal findings and conceptual model. ( A ) ACE2-Fc binding was measured to wild-type or glycoEnzyme-KO 293 T cells expressing Spike. Sialidase treatment of cells was performed in some cases. Similar studies also measured S1-Fc and RBD-Fc binding to cell-surface expressed ACE2. ( B ) SARS-CoV-2 pseudovirus (bearing Spike-WT, Spike-mut, Spike-delta variants) were generated in wild-type or glycoEnzyme-KO 293Ts, in the presence and absence of kifunensine. Main results of binding ( A ) and viral entry ( B ) assay are listed. ( C ) Conceptual model shows that kifunensine can induce S1-S2 site proteolysis on Spike-WT and Spike-mut virus, but not Spike-delta virus. This proteolysis reduces RBD presentation and attenuates viral entry into 293T/ACE2. Without affecting S1-S2 cleavage, kifunensine also partially reduced Spike-delta pseudovirus entry function. The data suggest additional roles for Spike N-glycans during viral entry.

    Techniques Used: Binding Assay, Expressing, Generated

    Sialidase treatment studies. ( A ) Sialidase protocol validation. All lectins were directly conjugated with Alexa dyes. They were incubated with cells at 1–5 µg/mL for 15 min before a quick wash and cytometry measurement. Compared to untreated control (left), sialidase treatment (right) decreased SNA lectin binding to α2,6 sialylated structures by 15-fold and increased ECL binding to desialylated lactosamine chains (Galβ1,4GlcNAcβ) by an order of magnitude. ( B ) Pseudovirus assay. DsRed fluorescence in HEK293T and stable 293T/ACE2 cells upon addition of VSVG, Spike-WT and Spike-mutant pseudotyped virus. ( C ) Sialidase treatment of pseudovirus. % DsRed positive cell data are shown for study in Figure 2F (main manuscript). Viral entry was sialidase independent. ( D ) Sialidase treatment of HEK/ACE2 cells. Pseudovirus expressing VSVG, Spike-WT and Spike-mutant were added to cells under conditions described in Figure 2G (main manuscript). All error bars are standard deviations. Data are representative of 3 independent runs.
    Figure Legend Snippet: Sialidase treatment studies. ( A ) Sialidase protocol validation. All lectins were directly conjugated with Alexa dyes. They were incubated with cells at 1–5 µg/mL for 15 min before a quick wash and cytometry measurement. Compared to untreated control (left), sialidase treatment (right) decreased SNA lectin binding to α2,6 sialylated structures by 15-fold and increased ECL binding to desialylated lactosamine chains (Galβ1,4GlcNAcβ) by an order of magnitude. ( B ) Pseudovirus assay. DsRed fluorescence in HEK293T and stable 293T/ACE2 cells upon addition of VSVG, Spike-WT and Spike-mutant pseudotyped virus. ( C ) Sialidase treatment of pseudovirus. % DsRed positive cell data are shown for study in Figure 2F (main manuscript). Viral entry was sialidase independent. ( D ) Sialidase treatment of HEK/ACE2 cells. Pseudovirus expressing VSVG, Spike-WT and Spike-mutant were added to cells under conditions described in Figure 2G (main manuscript). All error bars are standard deviations. Data are representative of 3 independent runs.

    Techniques Used: Incubation, Cytometry, Binding Assay, Fluorescence, Mutagenesis, Expressing

    Lectin binding to wild-type and glycogene-KO 293 T cells. A panel of lectins (from Vector Labs) was conjugated with Alexa dyes, either Alexa 405, 488 or 647. The binding of these fluorescent reagents to wild-type 293T, [N] - 293T and [O] - 293 T cells was measured using flow cytometry. The lectins bound: ( A ) N-glycan high-mannose and complex structures [ConA and LCA bind αMan in high-mannose glycans; PHA-L and PHA-E bind complex glycans], ( B ) lactosamine chains primarily on N-linked glycans [RCA, ECL bind terminal Gal or lactose; DSL bind β1,4GlcNAc], and ( C ) O-glycan related structures [PNA binds Galβ1,GalNAc; VVA and SBA bind GalNAcα]. Measurements were made with either untreated or sialidase treated 293 T cells. As seen: i. Knocking out MGAT1 in [N] - 293T reduces lectin binding in panels A and B (see arrow). ii. Knocking out C1GalT1 results in a dramatic decrease in PNA binding and increase in VVA and SBA binding, These data are consistent with the expected changes in lectin profile upon knocking out these N- and O-glycan-specific enzymes.
    Figure Legend Snippet: Lectin binding to wild-type and glycogene-KO 293 T cells. A panel of lectins (from Vector Labs) was conjugated with Alexa dyes, either Alexa 405, 488 or 647. The binding of these fluorescent reagents to wild-type 293T, [N] - 293T and [O] - 293 T cells was measured using flow cytometry. The lectins bound: ( A ) N-glycan high-mannose and complex structures [ConA and LCA bind αMan in high-mannose glycans; PHA-L and PHA-E bind complex glycans], ( B ) lactosamine chains primarily on N-linked glycans [RCA, ECL bind terminal Gal or lactose; DSL bind β1,4GlcNAc], and ( C ) O-glycan related structures [PNA binds Galβ1,GalNAc; VVA and SBA bind GalNAcα]. Measurements were made with either untreated or sialidase treated 293 T cells. As seen: i. Knocking out MGAT1 in [N] - 293T reduces lectin binding in panels A and B (see arrow). ii. Knocking out C1GalT1 results in a dramatic decrease in PNA binding and increase in VVA and SBA binding, These data are consistent with the expected changes in lectin profile upon knocking out these N- and O-glycan-specific enzymes.

    Techniques Used: Binding Assay, Plasmid Preparation, Flow Cytometry

    Related Articles

    Activity Assay:

    Article Title: Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration
    Article Snippet: .. Sialidase activity was verified based on SNA and ECL staining. ..

    Staining:

    Article Title: Inhibition of SARS-CoV-2 viral entry upon blocking N- and O-glycan elaboration
    Article Snippet: .. Sialidase activity was verified based on SNA and ECL staining. ..

    Purification:

    Article Title: FGF23 contains two distinct high-affinity binding sites enabling bivalent interactions with α-Klotho
    Article Snippet: .. Purified FGF23 variants were treated with O-glycosidase and α-(2→3,6,8,9)-neuraminidase (New England BioLabs) for 4 h at 37 °C as directed by manufacturer’s protocol. ..

    Article Title: Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
    Article Snippet: .. After washing the beads with PBS, the beads were incubated with or without the α2-3,6,8,9 Neuraminidase A (Sialidase; P0722; New England Biolabs) in a reaction solution, which is provided by the manufacturer, to remove sialic acid from purified NCAM1 proteins at 37 °C overnight. ..

    Modification:

    Article Title: Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
    Article Snippet: .. Removal of sialic acid modification on NCAM1Soluble or membrane fraction of HC and OB from BACE1+/+ and BACE1−/− mice was incubated with or without the α2-3,6,8,9 Neuraminidase A (Sialidase; P0722; New England Biolabs) in a reaction solution, which is provided by the manufacturer, to remove the sialic acid from NCAM1 proteins at 37 °C overnight. ..

    Mouse Assay:

    Article Title: Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
    Article Snippet: .. Removal of sialic acid modification on NCAM1Soluble or membrane fraction of HC and OB from BACE1+/+ and BACE1−/− mice was incubated with or without the α2-3,6,8,9 Neuraminidase A (Sialidase; P0722; New England Biolabs) in a reaction solution, which is provided by the manufacturer, to remove the sialic acid from NCAM1 proteins at 37 °C overnight. ..

    Incubation:

    Article Title: Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
    Article Snippet: .. Removal of sialic acid modification on NCAM1Soluble or membrane fraction of HC and OB from BACE1+/+ and BACE1−/− mice was incubated with or without the α2-3,6,8,9 Neuraminidase A (Sialidase; P0722; New England Biolabs) in a reaction solution, which is provided by the manufacturer, to remove the sialic acid from NCAM1 proteins at 37 °C overnight. ..

    Article Title: Spatiotemporal processing of neural cell adhesion molecules 1 and 2 by BACE1 in vivo
    Article Snippet: .. After washing the beads with PBS, the beads were incubated with or without the α2-3,6,8,9 Neuraminidase A (Sialidase; P0722; New England Biolabs) in a reaction solution, which is provided by the manufacturer, to remove sialic acid from purified NCAM1 proteins at 37 °C overnight. ..

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    New England Biolabs pngase f native
    Altered mobility of α-DG in COG-deficient CHO cells is a function of COG-dependent defects in O- glycosylation. A , representative Western blotting of WT CHO and ldlB cell lysates with or without <t>PNGase</t> F digestion ( n = 2). The cells were treated with or without V. cholerae neuraminidase (120 milliunits/ml) prior to lysis. B , cells were cultured with increasing concentrations of a furin inhibitor, and Western blotting analysis was performed on lysates using an anti-DG (core) antibody ( n = 2). Furin inhibition results in a comparable increase in the apparent molecular weight of α-DG in both WT CHO and ldlB cells. C , cells were treated with two different amounts (60 or 120 milliunits/ml) of V. cholerae neuraminidase, and Western blotting analysis was performed. D , Western blotting of α-DG in WT, ldlB, and cog1 -corrected ldlB cells treated with or without V. cholerae neuraminidase ( n = 2). IB , immunoblot.
    Pngase F Native, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    New England Biolabs α2 3
    HPLC-FLD profile of procainamide labeled N -glycans from rituximab and the glycoengineered rituximab with α2,6 linked sialic acid. A). HPLC profile of rituximab N -glycans with major species shown. B). The glycan profile from the glycoengineered rituximab prepared by Endo S digestion. The major glycan is G2FS2 with α2,6 linked sialic acid. C). The glycan profile from the glycoengineered rituximab after α2-3 Neuraminidase S treatment. D). The glycan profile from the glycoengineered rituximab after <t>α2-3,6,8,9</t> Neuraminidase A treatment. E). The glycan profile from the glycoengineered rituximab prepared by Endo S2 digestion.
    α2 3, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs α2 3 neuraminidase
    EICs of A2G2S2, A2G2S1 and A2G2 glycopeptides in non-treated (a-c) , <t>α2–3</t> neuraminidase treated (d-f) and α2–3,6,8,9 neuraminidase treated samples (g-i) . The peaks at 37.3 min and 39.2 min in (a) showed sensitivity for α2–3 neuraminidase, which disappeared after enzymatic digestion (d) , indicating these two peaks were containing at least one sialic acid that were α2–3 linked to galactose. Upon α2–3 neuraminidase digestion, A2G2S1 can be detected (e) , which were not originally in sample (b) . Since the 35.5 min peak in (a) remained after α2–3 neuraminidase digestion (d) , both the sialic acid linkages can be assigned α2–6. Comparing the intensities between (e) and (f) , the 37.3 min peak in (a) .
    α2 3 Neuraminidase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Altered mobility of α-DG in COG-deficient CHO cells is a function of COG-dependent defects in O- glycosylation. A , representative Western blotting of WT CHO and ldlB cell lysates with or without PNGase F digestion ( n = 2). The cells were treated with or without V. cholerae neuraminidase (120 milliunits/ml) prior to lysis. B , cells were cultured with increasing concentrations of a furin inhibitor, and Western blotting analysis was performed on lysates using an anti-DG (core) antibody ( n = 2). Furin inhibition results in a comparable increase in the apparent molecular weight of α-DG in both WT CHO and ldlB cells. C , cells were treated with two different amounts (60 or 120 milliunits/ml) of V. cholerae neuraminidase, and Western blotting analysis was performed. D , Western blotting of α-DG in WT, ldlB, and cog1 -corrected ldlB cells treated with or without V. cholerae neuraminidase ( n = 2). IB , immunoblot.

    Journal: The Journal of Biological Chemistry

    Article Title: Defective mucin-type glycosylation on α-dystroglycan in COG-deficient cells increases its susceptibility to bacterial proteases

    doi: 10.1074/jbc.RA118.003014

    Figure Lengend Snippet: Altered mobility of α-DG in COG-deficient CHO cells is a function of COG-dependent defects in O- glycosylation. A , representative Western blotting of WT CHO and ldlB cell lysates with or without PNGase F digestion ( n = 2). The cells were treated with or without V. cholerae neuraminidase (120 milliunits/ml) prior to lysis. B , cells were cultured with increasing concentrations of a furin inhibitor, and Western blotting analysis was performed on lysates using an anti-DG (core) antibody ( n = 2). Furin inhibition results in a comparable increase in the apparent molecular weight of α-DG in both WT CHO and ldlB cells. C , cells were treated with two different amounts (60 or 120 milliunits/ml) of V. cholerae neuraminidase, and Western blotting analysis was performed. D , Western blotting of α-DG in WT, ldlB, and cog1 -corrected ldlB cells treated with or without V. cholerae neuraminidase ( n = 2). IB , immunoblot.

    Article Snippet: A. ureafaciens neuraminidase (P0722) PNGase F (P0704S) were purchased from New England Biolabs.

    Techniques: Western Blot, Lysis, Cell Culture, Inhibition, Molecular Weight

    HPLC-FLD profile of procainamide labeled N -glycans from rituximab and the glycoengineered rituximab with α2,6 linked sialic acid. A). HPLC profile of rituximab N -glycans with major species shown. B). The glycan profile from the glycoengineered rituximab prepared by Endo S digestion. The major glycan is G2FS2 with α2,6 linked sialic acid. C). The glycan profile from the glycoengineered rituximab after α2-3 Neuraminidase S treatment. D). The glycan profile from the glycoengineered rituximab after α2-3,6,8,9 Neuraminidase A treatment. E). The glycan profile from the glycoengineered rituximab prepared by Endo S2 digestion.

    Journal: mAbs

    Article Title: Evaluation of a glycoengineered monoclonal antibody via LC-MS analysis in combination with multiple enzymatic digestion

    doi: 10.1080/19420862.2015.1113361

    Figure Lengend Snippet: HPLC-FLD profile of procainamide labeled N -glycans from rituximab and the glycoengineered rituximab with α2,6 linked sialic acid. A). HPLC profile of rituximab N -glycans with major species shown. B). The glycan profile from the glycoengineered rituximab prepared by Endo S digestion. The major glycan is G2FS2 with α2,6 linked sialic acid. C). The glycan profile from the glycoengineered rituximab after α2-3 Neuraminidase S treatment. D). The glycan profile from the glycoengineered rituximab after α2-3,6,8,9 Neuraminidase A treatment. E). The glycan profile from the glycoengineered rituximab prepared by Endo S2 digestion.

    Article Snippet: Trypsin-ultra (P8101S), PNGase F (P0708S), α2-3 Neuraminidase S (P0743S), α2-3,6,8,9 Neuraminidase A (P0722S), Remove-iT Endo S (P0741S) are products from New England Biolabs.

    Techniques: High Performance Liquid Chromatography, Labeling

    EICs of A2G2S2, A2G2S1 and A2G2 glycopeptides in non-treated (a-c) , α2–3 neuraminidase treated (d-f) and α2–3,6,8,9 neuraminidase treated samples (g-i) . The peaks at 37.3 min and 39.2 min in (a) showed sensitivity for α2–3 neuraminidase, which disappeared after enzymatic digestion (d) , indicating these two peaks were containing at least one sialic acid that were α2–3 linked to galactose. Upon α2–3 neuraminidase digestion, A2G2S1 can be detected (e) , which were not originally in sample (b) . Since the 35.5 min peak in (a) remained after α2–3 neuraminidase digestion (d) , both the sialic acid linkages can be assigned α2–6. Comparing the intensities between (e) and (f) , the 37.3 min peak in (a) .

    Journal: Analytical chemistry

    Article Title: Isomeric Separation of N-Glycopeptides Derived from Glycoproteins by Porous Graphitic Carbon (PGC) LC-MS/MS

    doi: 10.1021/acs.analchem.0c00668

    Figure Lengend Snippet: EICs of A2G2S2, A2G2S1 and A2G2 glycopeptides in non-treated (a-c) , α2–3 neuraminidase treated (d-f) and α2–3,6,8,9 neuraminidase treated samples (g-i) . The peaks at 37.3 min and 39.2 min in (a) showed sensitivity for α2–3 neuraminidase, which disappeared after enzymatic digestion (d) , indicating these two peaks were containing at least one sialic acid that were α2–3 linked to galactose. Upon α2–3 neuraminidase digestion, A2G2S1 can be detected (e) , which were not originally in sample (b) . Since the 35.5 min peak in (a) remained after α2–3 neuraminidase digestion (d) , both the sialic acid linkages can be assigned α2–6. Comparing the intensities between (e) and (f) , the 37.3 min peak in (a) .

    Article Snippet: Murine IgG1 (Intact mAb Mass Check Standard) was obtained from Waters Corporation (Milford, MA). α2–3 neuraminidase, α2–3,6,8,9 neuraminidase, β1–3 galactosidase, β1–4 galactosidase and β1–3,4 galactosidase were acquired from New England Biolabs (Ipswich, MA).

    Techniques:

    N -glycosylation changes in squamous cell carcinoma (SCC). (A) Supervised hierarchical cluster analysis of healthy and tumor skin. Rows display each of the 15 patient glycan data (T, tumor tissue; C, healthy control). Columns indicate the N -glycan ID. Five clusters can be observed: in healthy tissue N -glycans with α2-3 linked N -acetylneuraminic acid (NeuAc) appeared to be present in increased levels whereas α2-6-NeuAc and oligomannose N -glycan levels were higher in tumor tissue. (B) Statistical evaluation of sialylated and oligomannose N -glycans uncovered significant changes [ p ≤ 0.04, using a t -test, indicated by an asterisk (*)]. Oligomannose N -glycans were upregulated whereas α2-3 linked NeuAc carrying N -glycans were down regulated in SCC.

    Journal: Frontiers in Oncology

    Article Title: Alterations of the Human Skin N- and O-Glycome in Basal Cell Carcinoma and Squamous Cell Carcinoma

    doi: 10.3389/fonc.2018.00070

    Figure Lengend Snippet: N -glycosylation changes in squamous cell carcinoma (SCC). (A) Supervised hierarchical cluster analysis of healthy and tumor skin. Rows display each of the 15 patient glycan data (T, tumor tissue; C, healthy control). Columns indicate the N -glycan ID. Five clusters can be observed: in healthy tissue N -glycans with α2-3 linked N -acetylneuraminic acid (NeuAc) appeared to be present in increased levels whereas α2-6-NeuAc and oligomannose N -glycan levels were higher in tumor tissue. (B) Statistical evaluation of sialylated and oligomannose N -glycans uncovered significant changes [ p ≤ 0.04, using a t -test, indicated by an asterisk (*)]. Oligomannose N -glycans were upregulated whereas α2-3 linked NeuAc carrying N -glycans were down regulated in SCC.

    Article Snippet: The glycan sample was reconstituted in 10 μL 25 mM ammonium acetate buffer (pH 5.5) and 25 U of a α2-3-specific neuraminidase (NEB, Ipswitch, MA, USA) were added.

    Techniques:

    Healthy human skin N -glycome. (A) Bean diagram representing the 10 most abundant N -glycans determined from 14 patients. Green bars indicate individual data points, the black line represents the median, and the gray area depicts the data density. Columns indicate the glycan structures given by their glycan ID (Table S1 in Supplementary Material). (B) Relative N -glycan class abundances found in healthy skin biopsies. Sialylated and fucosylated structures were the major components representing the human skin N -glycome. (C) Relative abundances of different structure features found on sialylated N -glycans. Blue bars represent sialylated N -glycans with and without core fucose depending on their sialic acid linkage, showing that core fucosylation was a more abundant feature on N -glycans carrying one or two α2-3 linked N -acetylneuraminic acid (NeuAc) residues (if both, α2-3 and α2-6 linkages were present on one N- glycan, this N -glycan was considered in both linkage categories). Overall, α2-6-linked NeuAc was a slightly more abundant feature compared with α2-3-linked NeuAc. Most of the sialylated N -glycans carried two NeuAc residues. Tri-antennary species were below 3% and rather low abundant using porous graphitized carbon, while multiplexed capillary gel electrophoresis with laser induced fluorescence detection (xCGE-LIF) detected slightly higher levels of tri- and tetra-antennary N -glycans (see Supplementary file “xCGE-LIF quant.xlsx” in Supplementary Material).

    Journal: Frontiers in Oncology

    Article Title: Alterations of the Human Skin N- and O-Glycome in Basal Cell Carcinoma and Squamous Cell Carcinoma

    doi: 10.3389/fonc.2018.00070

    Figure Lengend Snippet: Healthy human skin N -glycome. (A) Bean diagram representing the 10 most abundant N -glycans determined from 14 patients. Green bars indicate individual data points, the black line represents the median, and the gray area depicts the data density. Columns indicate the glycan structures given by their glycan ID (Table S1 in Supplementary Material). (B) Relative N -glycan class abundances found in healthy skin biopsies. Sialylated and fucosylated structures were the major components representing the human skin N -glycome. (C) Relative abundances of different structure features found on sialylated N -glycans. Blue bars represent sialylated N -glycans with and without core fucose depending on their sialic acid linkage, showing that core fucosylation was a more abundant feature on N -glycans carrying one or two α2-3 linked N -acetylneuraminic acid (NeuAc) residues (if both, α2-3 and α2-6 linkages were present on one N- glycan, this N -glycan was considered in both linkage categories). Overall, α2-6-linked NeuAc was a slightly more abundant feature compared with α2-3-linked NeuAc. Most of the sialylated N -glycans carried two NeuAc residues. Tri-antennary species were below 3% and rather low abundant using porous graphitized carbon, while multiplexed capillary gel electrophoresis with laser induced fluorescence detection (xCGE-LIF) detected slightly higher levels of tri- and tetra-antennary N -glycans (see Supplementary file “xCGE-LIF quant.xlsx” in Supplementary Material).

    Article Snippet: The glycan sample was reconstituted in 10 μL 25 mM ammonium acetate buffer (pH 5.5) and 25 U of a α2-3-specific neuraminidase (NEB, Ipswitch, MA, USA) were added.

    Techniques: Nucleic Acid Electrophoresis, Fluorescence