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Label‐Free Quantitative Proteomics versus Antibody‐Based Assays to Measure Neutrophil‐Derived Enzymes in Saliva
Author(s) -
Silbereisen Angelika,
Alassiri Saeed,
Bao Kai,
Grossmann Jonas,
Nanni Paolo,
Fernandez Claudia,
Tervahartiala Taina,
Nascimento Gustavo G.,
Belibasakis Georgios N.,
Heikkinen AnnaMaria,
Lopez Rodrigo,
Sorsa Timo,
Bostanci Nagihan
Publication year - 2020
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201900050
Subject(s) - saliva , biomarker , matrix metalloproteinase , antibody , immunoassay , quantitative proteomics , elastase , myeloperoxidase , inflammation , immunology , gingivitis , medicine , microbiology and biotechnology , biology , proteomics , enzyme , biochemistry , dentistry , gene
Purpose This study aims to validate label‐free quantitative proteomics (LFQ) against antibody‐based methods for quantifying established periodontal disease biomarkers in saliva. Experimental Design In an experimental gingivitis model, healthy volunteers ( n = 10) provide saliva at baseline (d0), during the induction (d7, d14, d21) and resolution (d35) of gingival inflammation (total n = 50). Biomarker levels are analyzed by LFQ and time‐resolved immunofluorometric assay (IFMA) or enzyme‐linked immunosorbent assay (ELISA). Molecular matrix metalloproteinase (MMP)‐8 forms are assessed by Western blot (WB) analysis. Results LFQ detects significantly ( p < 0.05) elevated MMP‐8 (d21vsd7, d35vsd7) and tissue inhibitor of matrix metalloproteinases (TIMP)‐1 (d35vsd7). Latent MMP‐8 (70–80 kDa) is present (d0–d35), but not active MMP‐8 (50–60 kDa). LFQ and immunoassay data significantly correlate for MMP‐8 ( r = 0.36), myeloperoxidase ( r = 0.39), polymorphonuclear leukocyte elastase ( r = 0.33), and TIMP‐1 ( r = −0.24). Conclusion and Clinical Relevance LFQ can quantify enzyme levels in saliva, however lacks the ability to measure enzymatic activity. WB analysis reveals that MMP‐8 may not be activated during induction of gingival inflammation. Significant but weak correlations between IFMA or ELISA and LFQ suggest a limited capacity of available antibodies to reliably quantify salivary biomarkers for periodontal diseases. Novel “anti‐peptide” antibodies designed by newer targeted mass spectrometry‐based approaches can help to overcome these drawbacks.