Journal: Arthritis Research & Therapy

Article Title: Alteration of matrix metalloproteinase-3 O -glycan structure as a biomarker for disease activity of rheumatoid arthritis

doi: 10.1186/s13075-016-1013-2

Figure Lengend Snippet: Schematic overview of the antibody-overlay lectin microarray. A total of 45 kinds of lectins are spotted in triplicate on the array slide. Immunoprecipitation samples were applied to them, and glycoproteins bound to the lectins through their glycans. After washing, the lectins that bound to matrix metalloproteinase (MMP)-3 glycans were detected using anti-MMP-3-specific antibody

Article Snippet: For sialic acid digestion, IP samples were incubated with Glyko Sialidase A (PROzyme, Hayward, CA, USA) at 37 °C for 1 h before applying them to lectin microarrays (LecChip; GlycoTechnica, Yokohama, Japan).

Techniques: Microarray, Immunoprecipitation

Journal: Arthritis Research & Therapy

Article Title: Alteration of matrix metalloproteinase-3 O -glycan structure as a biomarker for disease activity of rheumatoid arthritis

doi: 10.1186/s13075-016-1013-2

Figure Lengend Snippet: The Agrocybe cylindracea galectin (ACG)/Jacalin index reflects rheumatoid arthritis (RA) activity. a Correlation of matrix metalloproteinase-3 (MMP-3) quantities before and after immunoprecipitation (IP). Serum MMP-3 measured by turbidimetric immunoassay and MMP-3 in IP samples measured by Western blot (WB) analysis ( n = 24). b Representative data derived from antibody-overlay lectin microarray. Data are from a patient with RA who had high disease activity and a patient with RA who was in remission. Net intensity from triplicate experiments is presented as the mean ± SD. c Correlation between immunoprecipitated MMP-3 quantity and lectin signal. Patients with more than moderate disease activity are represented by open symbols ; otherwise, data are shown as filled symbols . d Correlation between the ACG/Jacalin index and Disease Activity Score in 28 joints with erythrocyte sedimentation rate (DAS28-ESR). MAL_I Maackia amurensis leukoagglutinin I, SNA Sambucus nigra agglutinin, PHA(E) Phaseolus vulgaris erythroagglutinin, NPA Narcissus pseudonarcissus agglutinin, BPL Bauhinia purpurea lectin, ABA Agaricus bisporus agglutinin, LEL Lycopersicon esculentum lectin, STL Solanum tuberosum lectin, PNA peanut agglutinin, ACA Amaranthus caudatus agglutinin, MPA Maclura pomifera agglutinin, MAH Maackia amurensis hemagglutinin, WGA wheat germ agglutinin

Article Snippet: For sialic acid digestion, IP samples were incubated with Glyko Sialidase A (PROzyme, Hayward, CA, USA) at 37 °C for 1 h before applying them to lectin microarrays (LecChip; GlycoTechnica, Yokohama, Japan).

Techniques: Activity Assay, Immunoprecipitation, Western Blot, Derivative Assay, Microarray, Sedimentation

Journal: Arthritis Research & Therapy

Article Title: Alteration of matrix metalloproteinase-3 O -glycan structure as a biomarker for disease activity of rheumatoid arthritis

doi: 10.1186/s13075-016-1013-2

Figure Lengend Snippet: Matrix metalloproteinase-3 (MMP-3) with elevated Agrocybe cylindracea galectin (ACG)/Jacalin signals produced in local lesions reflects the degree of sialylation. a Synovial fluid MMP-3 was measured by Western blot analysis after immunoprecipitation. Patients with RA ( lanes 1–3 ) and patients with OA ( lanes 4–6 ). b Representative lectin signals derived from synovial MMP-3 (mean ± SD). c The structure of T and sialyl-T antigen. Jacalin recognizes the former. d Jacalin and ACG signals compared before and after sialic acid digestion (mean ± SD). e The estimated basic structure of MMP-3 O -glycan. ABA Agaricus bisporus agglutinin, UDA Urtica dioica agglutinin , ACA Amaranthus caudatus agglutinin, WGA wheat germ agglutinin

Article Snippet: For sialic acid digestion, IP samples were incubated with Glyko Sialidase A (PROzyme, Hayward, CA, USA) at 37 °C for 1 h before applying them to lectin microarrays (LecChip; GlycoTechnica, Yokohama, Japan).

Techniques: Produced, Western Blot, Immunoprecipitation, Derivative Assay, Glycoproteomics