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Evaluation and Validation of the Detection of soluble Triggering Receptor Expressed on Myeloid Cells 1 by Enzyme-linked immunosorbent Assay.

Hasibeder A, Stein P, Brandwijk R, Schild H, Radsak MP - Sci Rep (2015)

Bottom Line: In addition, a soluble TREM-1 variant (sTREM-1) is detectable in sera or bronchoalveolar-lavage fluids from patients.Investigating potential confounding factors for sTREM-1 detection, serum heat-inactivation (HI) showed improved recovery compared to non-HI (NHI) serum, reproducible by addition of complement and re-heat-inactivation.Immunoassays for research use only are in general hampered by lack of standardization.

View Article: PubMed Central - PubMed

Affiliation: IIIrd Department of Medicine, Johannes-Gutenberg University Medical Center, Mainz, Germany.

ABSTRACT
Triggering receptor expressed on myeloid cells (TREM)-1 plays an important role in innate immune responses and is upregulated under infectious as well as non-infectious conditions. In addition, a soluble TREM-1 variant (sTREM-1) is detectable in sera or bronchoalveolar-lavage fluids from patients. Currently, various studies are difficult to compare, since the methods of detection by enzyme-linked immunosorbent assays (ELISA) vary among different research groups. In this study, we compared three different s-TREM-1 specific ELISAs and identified individual assay characteristics finding notable differences in sTREM-1 concentrations in part depending on the employed buffers. Investigating potential confounding factors for sTREM-1 detection, serum heat-inactivation (HI) showed improved recovery compared to non-HI (NHI) serum, reproducible by addition of complement and re-heat-inactivation. Hence we identified complement as a heat-sensitive confounder in some sTREM-1 ELISAs. We conclude that it is difficult to directly compare data of several studies, in particular if different ELISAs are engaged. Immunoassays for research use only are in general hampered by lack of standardization. Further standardization is needed until sTREM-1 ELISA is capable for better reproducibility of studies and clinical application.

No MeSH data available.


Related in: MedlinePlus

Comparison of sTREM-1 specific ELISAs.Comparison of curve progression of rh-sTREM-1 standard in sTREM-1 Radsak (A), R&D (B), R&D + pcb (C) and Hycult Biotech (D) ELISA. 2,000 pg/mL of appropriate rh-sTREM-1 was employed and further twofold serially diluted for each ELISA. The OD was measured and depicted as the mean value of duplicates. One representative experiment out of three repeats is shown. The curves follow a four-parametric nonlinear curve fitting.
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f1: Comparison of sTREM-1 specific ELISAs.Comparison of curve progression of rh-sTREM-1 standard in sTREM-1 Radsak (A), R&D (B), R&D + pcb (C) and Hycult Biotech (D) ELISA. 2,000 pg/mL of appropriate rh-sTREM-1 was employed and further twofold serially diluted for each ELISA. The OD was measured and depicted as the mean value of duplicates. One representative experiment out of three repeats is shown. The curves follow a four-parametric nonlinear curve fitting.

Mentions: Figure 1 shows that all dilution series of the ELISA specific rh-sTREM-1 reveal a typical sigmoid curve course in the semi-log plot. The four-parametric nonlinear regression resulted in good correlation (R2 = 1) and comparable high slopes (Radsak 0.9602, R&D 1.011, R&D + pcb 1.060, Hycult Biotech 1.134) for all ELISAs.


Evaluation and Validation of the Detection of soluble Triggering Receptor Expressed on Myeloid Cells 1 by Enzyme-linked immunosorbent Assay.

Hasibeder A, Stein P, Brandwijk R, Schild H, Radsak MP - Sci Rep (2015)

Comparison of sTREM-1 specific ELISAs.Comparison of curve progression of rh-sTREM-1 standard in sTREM-1 Radsak (A), R&D (B), R&D + pcb (C) and Hycult Biotech (D) ELISA. 2,000 pg/mL of appropriate rh-sTREM-1 was employed and further twofold serially diluted for each ELISA. The OD was measured and depicted as the mean value of duplicates. One representative experiment out of three repeats is shown. The curves follow a four-parametric nonlinear curve fitting.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612298&req=5

f1: Comparison of sTREM-1 specific ELISAs.Comparison of curve progression of rh-sTREM-1 standard in sTREM-1 Radsak (A), R&D (B), R&D + pcb (C) and Hycult Biotech (D) ELISA. 2,000 pg/mL of appropriate rh-sTREM-1 was employed and further twofold serially diluted for each ELISA. The OD was measured and depicted as the mean value of duplicates. One representative experiment out of three repeats is shown. The curves follow a four-parametric nonlinear curve fitting.
Mentions: Figure 1 shows that all dilution series of the ELISA specific rh-sTREM-1 reveal a typical sigmoid curve course in the semi-log plot. The four-parametric nonlinear regression resulted in good correlation (R2 = 1) and comparable high slopes (Radsak 0.9602, R&D 1.011, R&D + pcb 1.060, Hycult Biotech 1.134) for all ELISAs.

Bottom Line: In addition, a soluble TREM-1 variant (sTREM-1) is detectable in sera or bronchoalveolar-lavage fluids from patients.Investigating potential confounding factors for sTREM-1 detection, serum heat-inactivation (HI) showed improved recovery compared to non-HI (NHI) serum, reproducible by addition of complement and re-heat-inactivation.Immunoassays for research use only are in general hampered by lack of standardization.

View Article: PubMed Central - PubMed

Affiliation: IIIrd Department of Medicine, Johannes-Gutenberg University Medical Center, Mainz, Germany.

ABSTRACT
Triggering receptor expressed on myeloid cells (TREM)-1 plays an important role in innate immune responses and is upregulated under infectious as well as non-infectious conditions. In addition, a soluble TREM-1 variant (sTREM-1) is detectable in sera or bronchoalveolar-lavage fluids from patients. Currently, various studies are difficult to compare, since the methods of detection by enzyme-linked immunosorbent assays (ELISA) vary among different research groups. In this study, we compared three different s-TREM-1 specific ELISAs and identified individual assay characteristics finding notable differences in sTREM-1 concentrations in part depending on the employed buffers. Investigating potential confounding factors for sTREM-1 detection, serum heat-inactivation (HI) showed improved recovery compared to non-HI (NHI) serum, reproducible by addition of complement and re-heat-inactivation. Hence we identified complement as a heat-sensitive confounder in some sTREM-1 ELISAs. We conclude that it is difficult to directly compare data of several studies, in particular if different ELISAs are engaged. Immunoassays for research use only are in general hampered by lack of standardization. Further standardization is needed until sTREM-1 ELISA is capable for better reproducibility of studies and clinical application.

No MeSH data available.


Related in: MedlinePlus