sba conjugated with fluorescein  (Vector Laboratories)

Journal: The Journal of Biological Chemistry

Article Title: Enzymatic tailoring of anionic glycans for characterizing lectin and antibody specificities in a microarray format

doi: 10.1016/j.jbc.2025.110915

Figure Lengend Snippet: Heat map displaying lectin binding to selected sialylated and sulfated structures . The heat map was generated using the GLAD tool and represents the relative fluorescent units (RFU) normalized to 70,000 as the maximum value. Relevant epitopes are shown, structures including either Neu5Ac or Neu5Gc as terminal sialic acid or different linkages are indicated with a slash. For complete structures, please refer to .

Article Snippet: Biotinylated lectins concanavalin A (ConA, 5 mg/ml), Erythrina cristagalli lectin (ECL, 5 mg/ml), Griffonia simplicifolia isolectin B4 (GSL-I, 0.5 mg/ml) and II (GSL-II, 2 mg/ml), Lens culinaris lectin (LCA, 5 mg/ml), Maackia amurensis lectin I and II (MAA-I, 2 mg/ml; MAA-II, 1 mg/ml), peanut lectin (PNA 5 mg/ml), Ricinus communis lectin I (RCA, 5 mg/ml), Sambucus nigra /Elderberry Bark lectin (SNA, 2 mg/ml), Vicia villosa lectin (VVA, 2 mg/ml), Wisteria floribunda agglutinin (WFA, 2 mg/ml) and wheat germ agglutinin (WGA, 5 mg/ml), as well as fluorescein-labeled soybean lectin (SBA, 2 mg/ml) were purchased from Vector Laboratories.

Techniques: Binding Assay, Generated

Journal: The Journal of Biological Chemistry

Article Title: Enzymatic tailoring of anionic glycans for characterizing lectin and antibody specificities in a microarray format

doi: 10.1016/j.jbc.2025.110915

Figure Lengend Snippet: Heat map displaying Gal-specific lectin binding to all structures . The heat map was generated using the GLAD tool and represents the relative fluorescent units (RFU) normalized to 70,000 as the maximum value. Terminal modifications (of either one or both antennae) are shown. Unmodified β4Gal is indicated as x (no modification) while free terminal β3Gal is marked (compounds 21 and 38 ). For complete structures please refer to .

Article Snippet: Biotinylated lectins concanavalin A (ConA, 5 mg/ml), Erythrina cristagalli lectin (ECL, 5 mg/ml), Griffonia simplicifolia isolectin B4 (GSL-I, 0.5 mg/ml) and II (GSL-II, 2 mg/ml), Lens culinaris lectin (LCA, 5 mg/ml), Maackia amurensis lectin I and II (MAA-I, 2 mg/ml; MAA-II, 1 mg/ml), peanut lectin (PNA 5 mg/ml), Ricinus communis lectin I (RCA, 5 mg/ml), Sambucus nigra /Elderberry Bark lectin (SNA, 2 mg/ml), Vicia villosa lectin (VVA, 2 mg/ml), Wisteria floribunda agglutinin (WFA, 2 mg/ml) and wheat germ agglutinin (WGA, 5 mg/ml), as well as fluorescein-labeled soybean lectin (SBA, 2 mg/ml) were purchased from Vector Laboratories.

Techniques: Binding Assay, Generated, Modification

Journal: Frontiers in Immunology

Article Title: Aberrant glycosylation patterns as potential biomarkers for diagnosis and disease progression in bullous pemphigoid

doi: 10.3389/fimmu.2025.1538126

Figure Lengend Snippet: Correlation analysis of serum and blister fluid glycosylation with BP clinical indicators, and diagnostic accuracy assessed by ROC analysis. (a) Evaluation of the correlation between 6 glycosylation structures (binding to PHA-L, SNA, PSA, SBA, RCA120, AAL) and clinical indicators (BP180 and BP230 antibody titers, eosinophil count and percentage, white blood cell count, total serum IgE titer, erythrocyte sedimentation rate, high-sensitivity C-reactive protein level, hospital stay duration, age) in serum. (b) Evaluation of the correlation between 6 glycosylation structures (binding to PHA-L, SNA, PSA, SBA, RCA120, AAL) and clinical indicators (BP180 and BP230 antibody titers, eosinophil count and percentage, total protein, albumin, sodium, potassium, calcium ion concentrations, age, and the six glycosylation modifications) in blister fluid. Spearman’s correlation analysis was used for statistical testing. X indicates p>0.05 after statistical testing. Red color represents positive correlation, while blue color represents negative correlation. The larger and deeper the circle, the stronger the correlation coefficient. (c) ROC analysis for serum glycopatterns recognized by lectin PSA, SNA, SBA to discriminate BP patients from healthy controls. (d) ROC analysis for blister fluid glycopatterns recognized by lectin RCA120, SBA, AAL to discriminate BP patients from control group. The area under the curve (AUC) is used as a metric of diagnostic ability.

Article Snippet: Briefly, during Lectin ELISA of serum and blister fluid, biotinylated lectins were diluted with 1% BSA to the following concentrations: HPA-L (Vector, B-1115) 5μg/mL, SNA (Vector, B-1305) 0.5μg/mL, PSA (Vector, B-1055) 0.5μg/mL, SBA (Vector, B-1015) 0.5μg/mL, RCA120 (Vector, B-1085) 0.05μg/mL, AAL (Vector, B-1395) 0.01μg/mL, while during Lectin ELISA of saliva, biotinylated lectins were diluted to the following concentrations: HPA-L 5μg/mL, SNA 2μg/mL, PSA 1μg/mL, SBA 5μg/mL, RCA120 1μg/mL, AAL 1μg/mL.

Techniques: Glycoproteomics, Diagnostic Assay, Binding Assay, Cell Counting, Sedimentation, Control

Journal: Frontiers in Immunology

Article Title: Aberrant glycosylation patterns as potential biomarkers for diagnosis and disease progression in bullous pemphigoid

doi: 10.3389/fimmu.2025.1538126

Figure Lengend Snippet: Alterations of glycopatterns in saliva of BP patients. (a) Comparative analysis of relative expression levels of 37 lectin-specific glycan structures in saliva samples of BP patients compared to healthy controls. The ratio of the BP group to the control group was logarithmically converted to a base of 1.5. A ratio >1 indicates an increase of more than 1.5 times compared to the control group, whereas a ratio <-1 denotes a decrease of over 0.67 times compared to the control group. High expression is depicted in red, while low expression is represented in blue. (b) Heat map of lectin with significant differentiation between BP patients and healthy controls. (c) The lectin ELISA confirmed that 3 glycans (binding to SNA, SBA, AAL) were significantly increased in the saliva of BP patients (n=20) compared to healthy controls (n=20). The data are presented as Mean ± SD. Analysis was performed using Wilcoxon Mann-Whitney test. (d) The ELISA was conducted to determine the positive rate of anti-BP180-NC16A antibodies in the saliva of BP patients. (e) Relationship between the presence of anti-BP180-NC16A antibodies in saliva and mucosal damage in BP patients. The data were presented as proportions, and statistical analysis was performed using chi-square test. (f) Differential analysis comparing the glycosylation structures (SNA) confirmed by lectin ELISA in the BP180 antibody-positive (n=12) and -negative groups (n=8) in saliva. The Wilcoxon Mann-Whitney test was used for statistical analysis.

Article Snippet: Briefly, during Lectin ELISA of serum and blister fluid, biotinylated lectins were diluted with 1% BSA to the following concentrations: HPA-L (Vector, B-1115) 5μg/mL, SNA (Vector, B-1305) 0.5μg/mL, PSA (Vector, B-1055) 0.5μg/mL, SBA (Vector, B-1015) 0.5μg/mL, RCA120 (Vector, B-1085) 0.05μg/mL, AAL (Vector, B-1395) 0.01μg/mL, while during Lectin ELISA of saliva, biotinylated lectins were diluted to the following concentrations: HPA-L 5μg/mL, SNA 2μg/mL, PSA 1μg/mL, SBA 5μg/mL, RCA120 1μg/mL, AAL 1μg/mL.

Techniques: Expressing, Glycoproteomics, Control, Enzyme-linked Immunosorbent Assay, Binding Assay, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Aberrant glycosylation patterns as potential biomarkers for diagnosis and disease progression in bullous pemphigoid

doi: 10.3389/fimmu.2025.1538126

Figure Lengend Snippet: Correlation of salivary glycosylation with BP clinical indicators and diagnostic accuracy of selected lectins via ROC analysis. (a) Correlation analysis was conducted to identify the relationship between changes in six glycan structures (specifically binding to PHA-L, SNA, PSA, SBA, RCA120, and AAL) in saliva and disease severity indicators (BP180 antibody titer, BP230 antibody titer, erythema and wheal score, mucosal damage score, BPDAI score). BP patients were categorized into four subgroups based on the positivity of salivary anti-BP180-NC16A and the presence of oral mucosal damage. Spearman’s correlation analysis was used for statistical testing. X indicates p>0.05 after statistical testing. Red color represents positive correlation, while blue color represents negative correlation. The larger and deeper the circle, the stronger the correlation coefficient. The numerical values in the middle represent the Spearman correlation coefficients. (b) Changes in the glycan structures bound to SNA, and AAL in the saliva of BP patients were detected using lectin ELISA before treatment at admission and after significant improvement at discharge (n=11). The Wilcoxon Signed-Rank test was used for statistical analysis. (c) ROC analysis was conducted to assess the glycan structures bound to SNA and AAL in saliva as biomarkers. AUC was used as an indicator to evaluate diagnostic capability.

Article Snippet: Briefly, during Lectin ELISA of serum and blister fluid, biotinylated lectins were diluted with 1% BSA to the following concentrations: HPA-L (Vector, B-1115) 5μg/mL, SNA (Vector, B-1305) 0.5μg/mL, PSA (Vector, B-1055) 0.5μg/mL, SBA (Vector, B-1015) 0.5μg/mL, RCA120 (Vector, B-1085) 0.05μg/mL, AAL (Vector, B-1395) 0.01μg/mL, while during Lectin ELISA of saliva, biotinylated lectins were diluted to the following concentrations: HPA-L 5μg/mL, SNA 2μg/mL, PSA 1μg/mL, SBA 5μg/mL, RCA120 1μg/mL, AAL 1μg/mL.

Techniques: Glycoproteomics, Diagnostic Assay, Binding Assay, Enzyme-linked Immunosorbent Assay