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Development of a newly immunoassay specific for mouse presepsin (sCD14-ST)
Gaku Takahashi1, Kouichi Hoshikawa2, Rioto Suzuki2
1Department of Critical Care, Disaster and General Medicine, School of Medicine, Iwate Medical University, 2-1-1 Idaidori Yahaba Town, Iwate, 028-3695, Japan. gakut@iwate-med.ac.jp.
Abstract:
Presepsin (sCD14-ST) is used as a marker for sepsis diagnosis. The production mechanism of presepsin is unique in that it is produced through phagocytosis of microorganisms. However, some studies have demonstrated that non-infected patients had increased presepsin levels and that presepsin is related to the risk or severity of diseases. This study was designed to describe a sensitive sandwich enzyme-linked immunosorbent assay for mouse presepsin developed to investigate the association of presepsin with diseases. Polyclonal antibodies were generated from peptide-immunized rabbit antiserum. Mouse presepsin standard was prepared using the recombinant method as an Fc-fusion protein. The linear detection range of the method was 4.7-300 pg/mL with a detection limit of 1.4 pg/mL. The assay detected mouse presepsin where mouse soluble CD14 (sCD14) was digested by cathepsin D proteinase and the cross-reactivity of sCD14 was not observed. The normal levels of mouse presepsin and sCD14 were compared; 65.9 ± 21.4 pg/mL and 43.2 ± 7.2 ng/mL were determined, respectively. Moreover, the levels of presepsin and sCD14 were compared with a lipopolysaccharide (LPS)-injected sepsis mouse model. The newly developed analytical method had high specificity to presepsin and is an efficient tool for studying the association between presepsin and diseases.
Insights
A new sensitive assay for mouse presepsin (sCD14-ST) was developed. This tool helps study presepsin
Area of Science:
- Biochemistry
- Immunology
- Biotechnology
Background:
- Presepsin (soluble CD14-sublyte) is a sepsis biomarker.
- Presepsin is produced via microorganism phagocytosis.
- Elevated presepsin levels occur in non-infected patients, suggesting broader disease relevance.
Purpose of the Study:
- Develop a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) for mouse presepsin.
- Investigate the association between presepsin and various diseases using the assay.
Main Methods:
- Generated polyclonal antibodies against mouse presepsin.
- Created a recombinant Fc-fusion protein standard for mouse presepsin.
- Validated the ELISA for sensitivity, specificity, and detection range (4.7-300 pg/mL).
Main Results:
- The assay demonstrated high specificity for presepsin, with no cross-reactivity to soluble CD14 (sCD14).
- Established normal levels for mouse presepsin (65.9 ± 21.4 pg/mL) and sCD14 (43.2 ± 7.2 ng/mL).
- Compared presepsin and sCD14 levels in a lipopolysaccharide-induced sepsis mouse model.
Conclusions:
- A highly specific and sensitive ELISA for mouse presepsin was successfully developed.
- This assay is an effective tool for researching presepsin's role in disease pathogenesis.
- Further studies can explore presepsin's association with disease risk and severity.
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