Characterization of binding and quantification of human autoantibodies to PDGFRα using a biosensor-based approach

Moroncini, Gianluca, Cuccioloni, Massimiliano, Mozzicafreddo, Matteo, Pozniak, Katarzyna Natalia, Grieco, Antonella, Paolini, Chiara, Tonnini, Cecilia, Spadoni, Tatiana, Svegliati, Silvia, Funaro, Ada, Angeletti, Mauro and Gabrielli, Armando (2017) Characterization of binding and quantification of human autoantibodies to PDGFRα using a biosensor-based approach. Analytical Biochemistry, 528 26-33. doi:10.1016/j.ab.2017.04.011

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Author Moroncini, Gianluca
Cuccioloni, Massimiliano
Mozzicafreddo, Matteo
Pozniak, Katarzyna Natalia
Grieco, Antonella
Paolini, Chiara
Tonnini, Cecilia
Spadoni, Tatiana
Svegliati, Silvia
Funaro, Ada
Angeletti, Mauro
Gabrielli, Armando
Title Characterization of binding and quantification of human autoantibodies to PDGFRα using a biosensor-based approach
Journal name Analytical Biochemistry   Check publisher's open access policy
ISSN 1096-0309
Publication date 2017-07-01
Sub-type Article (original research)
DOI 10.1016/j.ab.2017.04.011
Open Access Status File (Author Post-print)
Volume 528
Start page 26
End page 33
Total pages 8
Place of publication Philadelphia, PA, United States
Publisher Elsevier
Language eng
Subject 1304 Biophysics
1303 Biochemistry
1312 Molecular Biology
1307 Cell Biology
Abstract Systemic sclerosis (SSc) is a chronic autoimmune disease of the connective tissue. The variety and clinical relevance of autoantibodies in SSc patients have been extensively studied, eventually identifying agonistic autoantibodies targeting the platelet-derived growth factor receptor alpha (PDGFRα), and representing potential biomarkers for SSc. We used a resonant mirror biosensor to characterize the binding between surface-blocked PDGFRα and PDGFRα-specific recombinant human monoclonal autoantibodies (mAbs) produced by SSc B cells, and detect/quantify serum autoimmune IgG with binding characteristics similar to the mAbs. Kinetic data showed a conformation-specific, high-affinity interaction between PDGFRα and mAbs, with equilibrium dissociation constants in the low-to-high nanomolar range. When applied to total serum IgG, the assay discriminated between SSc patients and healthy controls, and allowed the rapid quantification of autoimmune IgG in the sera of SSc patients, with anti-PDGFRα IgG falling in the range 3.20–4.67 neq/L of SSc autoantibodies. The test was validated by comparison to direct and competitive anti-PDGFRα antibody ELISA. This biosensor assay showed higher sensibility with respect to ELISA, and other major advantages such as the specificity, rapidity, and reusability of the capturing surface, thus representing a feasible approach for the detection and quantification of high affinity, likely agonistic, SSc-specific anti-PDGFRα autoantibodies.
Keyword Autoantibody quantification
Binding kinetics
Systemic sclerosis
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
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