Journal article
Label-Free Quantitative Proteomics versus Antibody-Based Assays to Measure Neutrophil-Derived Enzymes in Saliva.
- Silbereisen A Section of Periodontology and Dental Prevention, Division of Oral Diseases, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels allé 8, 14152, Huddinge, Stockholm, Sweden.
- Alassiri S Department of Oral and Maxillofacial Diseases, University of Helsinki, Helsinki University Central Hospital, P.O. Box 41 (Mannerheimintie 172), 00014, Helsinki, Finland.
- Bao K Section of Periodontology and Dental Prevention, Division of Oral Diseases, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels allé 8, 14152, Huddinge, Stockholm, Sweden.
- Grossmann J Functional Genomics Center Zurich, ETH Zurich and University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
- Nanni P Functional Genomics Center Zurich, ETH Zurich and University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
- Fernandez C Functional Genomics Center Zurich, ETH Zurich and University of Zurich, Winterthurerstrasse 190, 8057, Zurich, Switzerland.
- Tervahartiala T Department of Oral and Maxillofacial Diseases, University of Helsinki, Helsinki University Central Hospital, P.O. Box 41 (Mannerheimintie 172), 00014, Helsinki, Finland.
- Nascimento GG Section of Periodontology, Department of Dentistry and Oral Health, Aarhus University, Vennelyst Boulevard 9, 8000, Aarhus C, Denmark.
- Belibasakis GN Section of Periodontology and Dental Prevention, Division of Oral Diseases, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels allé 8, 14152, Huddinge, Stockholm, Sweden.
- Heikkinen AM Department of Oral and Maxillofacial Diseases, University of Helsinki, Helsinki University Central Hospital, P.O. Box 41 (Mannerheimintie 172), 00014, Helsinki, Finland.
- Lopez R Section of Periodontology, Department of Dentistry and Oral Health, Aarhus University, Vennelyst Boulevard 9, 8000, Aarhus C, Denmark.
- Sorsa T Section of Periodontology and Dental Prevention, Division of Oral Diseases, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels allé 8, 14152, Huddinge, Stockholm, Sweden.
- Bostanci N Section of Periodontology and Dental Prevention, Division of Oral Diseases, Department of Dental Medicine, Karolinska Institutet, Alfred Nobels allé 8, 14152, Huddinge, Stockholm, Sweden.
- 2019-12-17
Published in:
- Proteomics. Clinical applications. - 2020
experimental gingivitis
periodontal disease
quantitative proteomics
saliva biomarkers
saliva proteases
English
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.
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.
- Language
-
- English
- Open access status
- closed
- Identifiers
-
- DOI 10.1002/prca.201900050
- PMID 31840410
- Persistent URL
- https://folia.unifr.ch/global/documents/193758
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