Journal article
Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
Archives of oral biology, Vol.57(2), pp.197-204
02/2012
DOI: 10.1016/j.archoralbio.2011.08.013
PMCID: PMC3260398
PMID: 21925642
Abstract
The short palate lung and nasal epithelial clone 1 (SPLUNC1) protein may be differentially expressed in oral infections, oral inflammatory disorders, or oral malignancies and may be involved in innate immune responses in the oral cavity. However, the actual concentration of SPLUNC1 in saliva has not previously been determined. In this study, we determined the concentrations of SPLUNC1 in saliva using a particle-based antibody capture and detection immunoassay. A commercial goat anti-rhSPLUNC1 polyclonal antibody (AF1897) was linked to fluorescent polystyrene microspheres and used as the capture antibody. A commercial mouse IgG2b anti-rhSPLUNC1 monoclonal antibody (MAB1897) was biotinylated and used as the detection antibody. Western blot and 2-dimensional fluorescence difference gel electrophoresis (2-D DIGE) analysis of immunoprecipitated rhSPLUNC1 and SPLUNC1 from saliva were used to show that the capture AF1897 and detection MAB1897 antibodies both recognized SPLUNC1. Protein concentrations in saliva from 20 subjects ranged from 0.9 to 23.9mg/ml; SPLUNC1 concentrations ranged from 34.7ng/ml to 13.8μg/ml; and SPLUNC concentrations normalized per mg of total salivary protein ranged from 4.7ng/ml to 5.3μg/ml. These results show that SPLUNC1 is detected in saliva in a variety of concentrations. This immunoassay may prove to be useful in determining the concentration of SPLUNC1 in saliva for assessing its role in the pathogenesis of oral infections, oral inflammatory disorders, or oral malignancies.
Details
- Title: Subtitle
- Quantitation of SPLUNC1 in saliva with an xMAP particle-based antibody capture and detection immunoassay
- Creators
- Karl G Kohlgraf - Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USAAbbey R Ackermann - Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USAKindra K Burnell - Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USARupasree N Srikantha - Department of Dermatology, Medical Laboratories, The University of Iowa, Iowa City, IA 52242, USASophie A Joly - Department of Infectious Diseases, Oakdale Research Park, The University of Iowa, Iowa City, IA 52242, USAJennifer A Bartlett - Department of Pediatrics, 240F EMRB, The University of Iowa, Iowa City, IA 52242, USALokesh Gakhar - Protein Crystallography Facility, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242, USAGeorgia K Johnson - Department of Periodontics, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USAPaul B McCray - Department of Pediatrics, 240F EMRB, The University of Iowa, Iowa City, IA 52242, USAJanet M Guthmiller - School of Dentistry, The University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAKim A Brogden - Dows Institute for Dental Research, College of Dentistry, The University of Iowa, Iowa City, IA 52242, USA
- Resource Type
- Journal article
- Publication Details
- Archives of oral biology, Vol.57(2), pp.197-204
- DOI
- 10.1016/j.archoralbio.2011.08.013
- PMID
- 21925642
- PMCID
- PMC3260398
- NLM abbreviation
- Arch Oral Biol
- ISSN
- 0003-9969
- eISSN
- 1879-1506
- Publisher
- Elsevier Ltd
- Language
- English
- Date published
- 02/2012
- Academic Unit
- Microbiology and Immunology; Pulmonary Medicine; Stead Family Department of Pediatrics; Biochemistry and Molecular Biology; Medicine Administration; Periodontics; Internal Medicine
- Record Identifier
- 9984065810302771
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