title: Development of a novel multiplexed assay for quantification of transforming growth factor-β (TGF-β).
abstract: Changes in activity or levels of transforming growth factor-β (TGF-β) are associated with a variety of diseases; however, measurement of TGF-β in biological fluids is highly variable. TGF-β is biologically inert when associated with its latency-associated peptide (LAP). Most available immunoassays require exogenous activation by acid/heat to release TGF-β from the latent complex. We developed a novel electrochemiluminescence-based multiplexed assay on the MesoScale Discovery® platform that eliminates artificial activation, simultaneously measures both active TGF-β1 and LAP1 and includes an internal control for platelet-derived TGF-β contamination in blood specimens. We optimized this assay to evaluate plasma levels as a function of activation type and clinical specimen preparation. We determined that breast cancer patients' plasma have higher levels of circulating latent TGF-β (LTGF-β) as measured by LAP1 than healthy volunteers (p < 0.0001). This assay provides a robust tool for correlative studies of LTGF-β levels with disease, treatment outcomes and toxicity with a broad clinical applicability.
authors.affiliation: Department of Radiation Oncology and.
authors.forename: Ilenia
authors.initials: I
authors.lastname: Pellicciotta
authors.forename: Ariel E
authors.initials: AE
authors.lastname: Marciscano
authors.forename: Matthew E
authors.initials: ME
authors.lastname: Hardee
authors.forename: Derek
authors.initials: D
authors.lastname: Francis
authors.forename: Silvia
authors.initials: S
authors.lastname: Formenti
authors.forename: Mary Helen
authors.initials: MH
authors.lastname: Barcellos-Hoff
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000097:blood*; D045744:Cell Line, Tumor; D017077:Culture Media, Conditioned; D004563:Electrochemistry / Q000379:methods; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D050883:HSC70 Heat-Shock Proteins / Q000737:chemistry; D064368:Healthy Volunteers; D006801:Humans; D007118:Immunoassay / Q000379:methods; D057230:Limit of Detection; D049449:Luminescence; D008163:Luminescent Measurements / Q000379:methods*; D008168:Lung / Q000166:cytology; D008907:Mink; D015203:Reproducibility of Results; D053773:Transforming Growth Factor beta1 / Q000737:chemistry*
chemical_list: D017077:Culture Media, Conditioned; D050883:HSC70 Heat-Shock Proteins; C494371:HSPA8 protein, human; C508960:TGFB1 protein, human; D053773:Transforming Growth Factor beta1
keywords: Assay development; LAP1; TGF-β; biological markers; plasma; radiation therapy