Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Circulating plasma antibody reactivity to oral commensals identified in pancreatic tumor lesions. Plasma diluent (1/300) from patients with histology verified pancreas tissues: IPMN with low grade dysplasia (LGD-IPMN), IPMN with high grade dysplasia or with invasive cancer (HGD-IPMN + Cancer), or benign tumors/non-IPMN (Control) was tested in duplicates in ELISA plates coated with indicated antigen in (A–F) (bacteria: 5 × 10E7 CFU/ml, peptide 10 μg/mL). The total plasma IgG level is shown in (G) . Internal control Mean OD absorbance values of duplicates are shown. Statistical analysis was performed using the Kolmogorov–Smirnov test, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Control, Enzyme-linked Immunosorbent Assay, Bacteria

Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Salivary IgA antibody reactivity to oral commensals quantified by ELISA using saliva diluent (1/16) from indicated patient group. Salivary IgA reactivity to F. nucleatum , the Fap2 mimotope, and S. gordonii (A,B,F) in HG-IPMN/Cancer patients is significantly increased compared to other groups. The differences in antibody levels against bacterial antigens of G. adiacens, S. anginosus , and E. coli (C,D,E) were insignificant between groups. Total plasma IgA level did not differ between the groups (G) . Statistical analysis was performed using the Kolmogorov–Smirnov test, * p < 0.05 and ** p < 0.01.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics

Journal: Frontiers in Immunology

Article Title: Circulating and Salivary Antibodies to Fusobacterium nucleatum Are Associated With Cystic Pancreatic Neoplasm Malignancy

doi: 10.3389/fimmu.2020.02003

Figure Lengend Snippet: Correlation between circulating plasma (A) or salivary (B) antibody reactivity to F. nucleatum and Fap2 mimotope, or between circulating plasma antibody and salivary antibodies to either of antigen (C,D) . Competitive ELISA (E) with saliva samples with or without antigen pre-absorption as indicated for 2 h, then subjected to Fap2 mimotope ELISA test. Statistical analysis was performed using two-tailed Pearson correlation test and Wilcoxon test for two related samples. ** p < 0.01, ns = not significant.

Article Snippet: Total levels of salivary IgA and plasma IgG antibodies were measured using Human IgA/IgG ELISA Kit (Novus Biologicals, Colorado, United States) according to the manufacturer’s instructions.

Techniques: Clinical Proteomics, Competitive ELISA, Enzyme-linked Immunosorbent Assay, Two Tailed Test

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: Current-potential profiles ( a , c ) and calibration curves ( b , d ) of neutralizing antibodies sIgA and IgG were obtained using the GLEIA−based electrochemical immunosensor. Current−potential responses were tested under different concentrations of ( a ) sIgA 100 ng/mL (blue), 20 ng/mL (grey), 5 ng/mL (orange), 1 ng/mL (red), and 0 ng/mL (black) and ( c ) IgG 62.5 U/mL (blue), 25 U/mL (grey), 6.25 U/mL (orange), 2.5 U/mL (red), and 0 U/mL (black). Calibration curves were indicated for sIgA ( b ) and IgG ( d ) as Michaelis–Menten-type functions by a non-linear curve fitted in the graphing software Origin2022 (OriginLab).

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Software

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: Comparisons of concentrations of neutralizing antibody IgG ( a ) and sIgA ( b ), antioxidant activity (indicated by luminescence inhibition rate) ( c ), and protein concentration ( d ) using saliva samples from 10 individuals are presented together. All 10 saliva samples were collected 3 weeks after the second dose of the vaccine.

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Antioxidant Activity Assay, Inhibition, Protein Concentration

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: The four datasets of neutralizing antibodies—IgG and sIgA, antioxidant activity, and protein concentration—measured in 10 samples are depicted individually in a radar chart for each individual. The numbers T1–T10 correspond to the sample numbers 1–10 in . The data are shown as a ratio of the maximum concentration of each measured item.

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Antioxidant Activity Assay, Protein Concentration, Concentration Assay

Journal: Biosensors

Article Title: Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva

doi: 10.3390/bios13020167

Figure Lengend Snippet: The results of continuous monitoring of the concentration of neutralizing IgG ( B ) and sIgA ( C ) antibodies, antioxidant activity ( D ), and protein concentration ( E ) in saliva and neutralizing IgG concentration in serum ( A ) before and after 1st, 2nd, and 3rd vaccinations in the same individual over time were compared vertically. The abscissa represents the date and time of sampling. 0: at first vaccination; 1a: 4 days later, 1b: 1 week later, 1c: 2 weeks later, and 1d: 3 weeks later; 2a: 4 days after second vaccination; 2b: 1 week later, 2c: 2 weeks later, 2d: 3 weeks later, 2e: 1 month later, 2f: 2 months later, 2g: 3 months later, 2h: 4 months later, 2i: 5 months later, 2j: 6 months later, and 2k: 8 months later; 3a: 3 days after the third vaccination, 3b: 2 weeks later, and 3c: 1 month later. The units on the vertical axis are as follows: ( A , B ) U/mL, ( C ) ng/mL, ( D ) %, and ( E ) mg/mL.

Article Snippet: As positive controls for neutralizing antibodies, the sIgA antibody (E-AB-V1027, Elabscience, Houston, TX, USA), IgG antibody (SPD-M180, Acro Biosystems, Tokyo, Japan), and IgG standard in the IgG ELISA kit (290-84201, FUJIFILM Wako, Osaka, Japan) were used.

Techniques: Concentration Assay, Antioxidant Activity Assay, Protein Concentration, Sampling

Journal:

Article Title: Cloning and characterization of an immunoglobulin A Fc receptor from cattle

doi: 10.1111/j.0019-2805.2003.01808.x

Figure Lengend Snippet: Ligand specificity of the bovine immunoglobulin A (IgA) Fc receptor (bFcαR). (a) COS-1 cells, transfected with either bFcαR or CD89 (without green fluorescent protein), were incubated with heat-aggregated (HA) human IgA preparations (HAhIgA), as indicated. Following a second incubation with the relevant anti-IgA–fluorescein isothiocyanate (FITC) conjugates, the cells were analysed on a FACScan flow cytometer. Results are representative of at least three separate experiments. PBS, phosphate-buffered saline.

Article Snippet: Sheep anti-bovine IgA-conjugated fluorescein isothiocyanate (FITC) was purchased from Serotec (Oxford, UK).

Techniques: Transfection, Incubation, Flow Cytometry, Saline

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Recombinant human IgA1 and IgA2 autoantibodies to type VII collagen induce subepidermal blistering ex vivo.

doi: 10.4049/jimmunol.1400160

Figure Lengend Snippet: FIGURE 1. Recombinant autoantibodies demonstrated the correct m.w. under reducing and nonreducing conditions, labeled the DEJ and bound to type VII collagen extracted from human dermis. (A) Recombinant autoantibodies were separated by 12% SDS-PAGE under reducing conditions and stained with Coomassie blue. The H chains of the recombinant proteins migrated at 50 (rIgG1), 60 (rIgA1), and 55 kDa (rIgA2), and the k L chains migrated at 25 kDa. The m.w. standard is indicated on the left. (B) Equal amounts of recombinant autoantibodies were separated by 7% SDS-PAGE under nonreducing conditions. The majority of the recombinant autoantibodies were assembled as complete 2H2L heterotetramers. In addition, for rIgA1 and rIgA2, additional bands at higher m.w. were detectable, indicating polymerization and a reduced amount of monomeric rIgA1 or rIgA2. (C) Immunoblotting of recombinant autoantibodies was performed using the same conditions as in (B), as well as detection with peroxidase-conjugated polyclonal anti-human k L chain Ab. The Coomassie staining of rIgA1 and rIgA2 appeared to be weaker overall than that for rIgG1, which may be because of the higher glycosylation content of IgA isotypes. The m.w. standard is indicated on the left. (D) IgA1 and IgA2 recombinant autoantibodies reacted with a 290-kDa band after immunoblotting of dermal extracts, whereas the isotype controls for IgA1 (Iso IgA1) and IgA2 (Iso IgA2) were negative. (E) Human neonatal foreskin cryosections were incubated with the different recombinant autoantibodies and visualized by isotype- and subclass-specific FITC-conjugated Abs. Both rIgA1 and rIgA2 presented a defined reactivity with the DEJ (arrows). As controls, a V-gene matched rIgG1 (31) and serum from a healthy volunteer (NHS) in conjunction with FITC-conjugated polyclonal rabbit anti-human IgG was used. Original magnification 3100.

Article Snippet: For the detection of Abs in cryosections, FITC-conjugated polyclonal goat anti-human IgA (Bio-Rad Laboratories GmbH, Munich, Germany) and polyclonal goat anti-human IgG (Sigma-Aldrich Chemie GmbH, Munich, Germany) were used as secondary Abs.

Techniques: Recombinant, Labeling, SDS Page, Staining, Western Blot, Glycoproteomics, Incubation