Journal: Journal of Immunology Research

Article Title: Epitope-Based Chicken-Derived Novel Anti-PAD2 Monoclonal Antibodies Inhibit Citrullination

doi: 10.1155/2021/6659960

Figure Lengend Snippet: PAD2 ribbon structure (PDB ID: 4N2C). The epitope 341-YLNRGDRWIQDEIEFGY-357 is shown in red, while calcium is shown in yellow. The three-dimensional structure of PAD2 was edited and prepared using UCSF chimera as described previously .

Article Snippet: Recombinant human PAD2 (rhPAD2) was purchased from Cayman Chemical (Michigan, USA) and was used for all experiments unless otherwise stated.

Techniques:

Journal: Journal of Immunology Research

Article Title: Epitope-Based Chicken-Derived Novel Anti-PAD2 Monoclonal Antibodies Inhibit Citrullination

doi: 10.1155/2021/6659960

Figure Lengend Snippet: Crossreactivity of chimeric anti-PAD2 antibodies with recombinant mouse PAD2. The values in the bracket indicates EC 50 values of antibodies against rhPAD2 and rmPAD2 enzymes. The enzyme-linked immunosorbent assay protocol is described .

Article Snippet: Recombinant human PAD2 (rhPAD2) was purchased from Cayman Chemical (Michigan, USA) and was used for all experiments unless otherwise stated.

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Journal of Immunology Research

Article Title: Epitope-Based Chicken-Derived Novel Anti-PAD2 Monoclonal Antibodies Inhibit Citrullination

doi: 10.1155/2021/6659960

Figure Lengend Snippet: Kinetics of binding of chimeric anti-PAD2 monoclonal antibodies (mAbs) with recombinant human PAD2.

Article Snippet: Recombinant human PAD2 (rhPAD2) was purchased from Cayman Chemical (Michigan, USA) and was used for all experiments unless otherwise stated.

Techniques: Binding Assay, Bioprocessing, Recombinant

Journal: Journal of Immunology Research

Article Title: Epitope-Based Chicken-Derived Novel Anti-PAD2 Monoclonal Antibodies Inhibit Citrullination

doi: 10.1155/2021/6659960

Figure Lengend Snippet: Ability of anti-PAD2 antibodies to inhibit PAD2-mediated citrullination of N α -benzoyl-L-arginine ethyl ester (BAEE). S4, S10, S24, S108, S170, and S309 antibodies were incubated with recombinant human PAD2 (rhPAD2) and buffer (20 mM Tris-HCl (pH 7.6), 150 mM NaCl, and 1 mM dithiothreitol) in a total volume of 44 μ L for 30 min at 25°C. BAEE and CaCl 2 (volume: 6 μ L) were added simultaneously and the mixture was stirred well. The mixture comprised 5 nM rhPAD2, 10 mM BAEE, 1 mM CaCl 2 , and various concentrations of antibodies (0.1–600 nM). The mixture was incubated at 37°C for 4 h, and 12.5 μ L of 5 M perchloric acid was added to stop the reaction. The level of citrullinated BAEE was determined using a colorimetric assay system.

Article Snippet: Recombinant human PAD2 (rhPAD2) was purchased from Cayman Chemical (Michigan, USA) and was used for all experiments unless otherwise stated.

Techniques: Incubation, Recombinant, Colorimetric Assay

Journal: Journal of Immunology Research

Article Title: Epitope-Based Chicken-Derived Novel Anti-PAD2 Monoclonal Antibodies Inhibit Citrullination

doi: 10.1155/2021/6659960

Figure Lengend Snippet: Ability of anti-PAD2 antibodies to inhibit recombinant human PAD2- (rhPAD2-) mediated histone citrullination. The anti-PAD2 antibodies were mixed with rhPAD2 and buffer as described in the legend of at 25°C for 30 min. Histone (1.8 mg/mL) and CaCl 2 (1 mM) were added simultaneously and the mixture was incubated for 4 h at 37°C. Citrullinated histone was determined using a colorimetric assay.

Article Snippet: Recombinant human PAD2 (rhPAD2) was purchased from Cayman Chemical (Michigan, USA) and was used for all experiments unless otherwise stated.

Techniques: Recombinant, Incubation, Colorimetric Assay

Journal: Journal of Immunology Research

Article Title: Epitope-Based Chicken-Derived Novel Anti-PAD2 Monoclonal Antibodies Inhibit Citrullination

doi: 10.1155/2021/6659960

Figure Lengend Snippet: Alanine scanning of epitopes along with their half-maximal effective concentration. EC 50 (ng/mL) values determined using enzyme-linked immunosorbent assay ∗ .

Article Snippet: Recombinant human PAD2 (rhPAD2) was purchased from Cayman Chemical (Michigan, USA) and was used for all experiments unless otherwise stated.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay, Residue