Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jan 18:10:1280681.
doi: 10.3389/fmolb.2023.1280681. eCollection 2023.

Development and application of an amplified luminescent proximity homogeneous assay-linked immunosorbent assay for the accurate quantification of kidney injury molecule-1

Yulin Fu  1 , Danqin Sun  2 , Yuan Qin  1 , Tianyu Zheng  1 , Zixuan Zhou  1 , Xiumei Zhou  1 , Xueqin Zhao  1 , Yan Xu  3 , Biao Huang  1
Affiliations

Development and application of an amplified luminescent proximity homogeneous assay-linked immunosorbent assay for the accurate quantification of kidney injury molecule-1

Yulin Fu et al. Front Mol Biosci. .

Abstract

Background: Kidney injury molecule-1 (Kim-1), a specific marker of kidney injury, is usually not expressed in normal kidneys or at very low levels but is highly expressed in injured renal tubular epithelial cells until the damaged cells recover completely. Therefore, we aimed to develop an efficient and highly sensitive assay to accurately quantify Kim-1 levels in human serum and urine. Methods: In this study, a novel immunoassay was developed and named amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA). Anti-Kim-1 antibodies can be directly coupled to carboxyl-modified donor and acceptor beads for the rapid detection of Kim-1 by double-antibody sandwich method. Serum and urine samples for Kim-1 measurements were obtained from 129 patients with nephropathy and 17 healthy individuals. Results: The linear range of Kim-1 detected by AlphaLISA was 3.83-5000 pg/mL, the coefficients of variation of intra-assay and inter-assay batches were 3.36%-4.71% and 5.61%-11.84%, respectively, and the recovery rate was 92.31%-99.58%. No cross reactions with neutrophil gelatinase-associated lipocalin, liver-type fatty acid binding protein, and matrix metalloproteinase-3 were observed. A good correlation (R 2 = 0.9086) was found between the findings of Kim-1-TRFIA and Kim-AlphaLISA for the same set of samples. In clinical trials, both serum and urine Kim-1 levels were significantly higher in patients with nephropathy than in healthy individuals, especially in patients with acute kidney injury. Furthermore, serum Kim-1 was superior to urinary Kim-1 in distinguishing between patients with nephropathy and healthy individuals. Conclusion: The developed Kim-1-AlphaLISA is highly efficient, precise, and sensitive, and it is suitable for the rapid detection of patients with acute kidney injury.

Keywords: acute kidney injury; amplified luminescent proximity homogeneous assay linked immunosorbent assay; double-antibody sandwich; kidney injury molecule-1; serum; urine.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Principle of AlphaLISA for detecting kidney injury molecule-1.
FIGURE 2
FIGURE 2
Selection of optimal donor bead concentration.
FIGURE 3
FIGURE 3
Selection of optimal receptor bead concentration.
FIGURE 4
FIGURE 4
Selection of optimal incubation time.
FIGURE 5
FIGURE 5
Standard curve of Kim-1.
FIGURE 6
FIGURE 6
Correlation analysis between the Kim-1-AlphaLISA and Kim-1-TRFIA.
FIGURE 7
FIGURE 7
(A) Serum Kim-1 concentrations in nephropathic patients and controls. (B) Urinary Kim-1 concentrations in patients with nephropathy and controls.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Armstrong C. M., Ruth L. E., Capobianco J. A., Strobaugh T. P., Rubio F. M., Gehring A. G. (2018). Detection of shiga toxin 2 produced by Escherichia coli in foods using a novel AlphaLISA. Toxins (Basel) 10 (11), 422. 10.3390/toxins10110422 - DOI - PMC - PubMed
    1. Bailly V., Zhang Z., Meier W., Cate R., Sanicola M., Bonventre J. V. (2002). Shedding of kidney injury molecule-1, a putative adhesion protein involved in renal regeneration. J. Biol. Chem. 277 (42), 39739–39748. 10.1074/jbc.M200562200 - DOI - PubMed
    1. Bellomo R., Kellum J. A., Ronco C. (2012). Acute kidney injury. Lancet 380 (9843), 756–766. 10.1016/S0140-6736(11)61454-2 - DOI - PubMed
    1. Campbell G. A., Hu D., Okusa M. D. (2014). Acute kidney injury in the cancer patient. Adv. Chronic Kidney Dis. 21 (1), 64–71. 10.1053/j.ackd.2013.08.002 - DOI - PubMed
    1. Chaturvedi S., Farmer T., Kapke G. F. (2009). Assay validation for KIM-1: human urinary renal dysfunction biomarker. Int. J. Biol. Sci. 5 (2), 128–134. 10.7150/ijbs.5.128 - DOI - PMC - PubMed

LinkOut - more resources