Highly sensitive reversed-phase high-performance liquid chromatography assay for the detection of Tamm–Horsfall protein in human urine

Masaru Akimoto, Eisaku Hokazono, Eri Ota, Takiko Tateishi, Yuzo Kayamori

研究成果: ジャーナルへの寄稿記事

抄録

Background: Tamm–Horsfall protein (also known as uromodulin) is the most abundant urinary protein in healthy individuals. Since initially characterized by Tamm and Horsfall, the amount of urinary excretion and structural mutations of Tamm–Horsfall protein is associated with kidney diseases. However, currently available assays for Tamm–Horsfall protein, which are mainly enzyme-linked immunosorbent assay-based, suffer from poor reproducibility and might give false negative results. Methods: We developed a novel, quantitative assay for Tamm–Horsfall protein using reversed-phase high-performance liquid chromatography. A precipitation pretreatment avoided urine matrix interference and excessive sample dilution. High-performance liquid chromatography optimization based on polarity allowed excellent separation of Tamm–Horsfall protein from other major urine components. Results: Our method exhibited high precision (based on the relative standard deviations of intraday [≤2.77%] and interday [≤5.35%] repetitions). The Tamm–Horsfall protein recovery rate was 100.0–104.2%. The mean Tamm–Horsfall protein concentration in 25 healthy individuals was 31.6 ± 18.8 mg/g creatinine. There was a strong correlation between data obtained by high-performance liquid chromatography and enzyme-linked immunosorbent assay (r = 0.906), but enzyme-linked immunosorbent assay values tended to be lower than high-performance liquid chromatography values at low Tamm–Horsfall protein concentrations. Conclusions: The high sensitivity and reproducibility of our Tamm–Horsfall protein assay will reduce the number of false negative results of the sample compared with enzyme-linked immunosorbent assay. Moreover, our method is superior to other high-performance liquid chromatography methods, and a simple protocol will facilitate further research on the physiological role of Tamm–Horsfall protein.

元の言語英語
ページ(範囲)75-84
ページ数10
ジャーナルAnnals of Clinical Biochemistry
53
発行部数1
DOI
出版物ステータス出版済み - 1 1 2016

Fingerprint

High performance liquid chromatography
Reverse-Phase Chromatography
Assays
High Pressure Liquid Chromatography
Urine
Immunosorbents
Proteins
Enzyme-Linked Immunosorbent Assay
Enzymes
Uromodulin
Kidney Diseases
Dilution
Creatinine
Recovery
Mutation

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

これを引用

Highly sensitive reversed-phase high-performance liquid chromatography assay for the detection of Tamm–Horsfall protein in human urine. / Akimoto, Masaru; Hokazono, Eisaku; Ota, Eri; Tateishi, Takiko; Kayamori, Yuzo.

:: Annals of Clinical Biochemistry, 巻 53, 番号 1, 01.01.2016, p. 75-84.

研究成果: ジャーナルへの寄稿記事

@article{6e8ec516c176422f8405259460f0d115,
title = "Highly sensitive reversed-phase high-performance liquid chromatography assay for the detection of Tamm–Horsfall protein in human urine",
abstract = "Background: Tamm–Horsfall protein (also known as uromodulin) is the most abundant urinary protein in healthy individuals. Since initially characterized by Tamm and Horsfall, the amount of urinary excretion and structural mutations of Tamm–Horsfall protein is associated with kidney diseases. However, currently available assays for Tamm–Horsfall protein, which are mainly enzyme-linked immunosorbent assay-based, suffer from poor reproducibility and might give false negative results. Methods: We developed a novel, quantitative assay for Tamm–Horsfall protein using reversed-phase high-performance liquid chromatography. A precipitation pretreatment avoided urine matrix interference and excessive sample dilution. High-performance liquid chromatography optimization based on polarity allowed excellent separation of Tamm–Horsfall protein from other major urine components. Results: Our method exhibited high precision (based on the relative standard deviations of intraday [≤2.77{\%}] and interday [≤5.35{\%}] repetitions). The Tamm–Horsfall protein recovery rate was 100.0–104.2{\%}. The mean Tamm–Horsfall protein concentration in 25 healthy individuals was 31.6 ± 18.8 mg/g creatinine. There was a strong correlation between data obtained by high-performance liquid chromatography and enzyme-linked immunosorbent assay (r = 0.906), but enzyme-linked immunosorbent assay values tended to be lower than high-performance liquid chromatography values at low Tamm–Horsfall protein concentrations. Conclusions: The high sensitivity and reproducibility of our Tamm–Horsfall protein assay will reduce the number of false negative results of the sample compared with enzyme-linked immunosorbent assay. Moreover, our method is superior to other high-performance liquid chromatography methods, and a simple protocol will facilitate further research on the physiological role of Tamm–Horsfall protein.",
author = "Masaru Akimoto and Eisaku Hokazono and Eri Ota and Takiko Tateishi and Yuzo Kayamori",
year = "2016",
month = "1",
day = "1",
doi = "10.1177/0004563215583698",
language = "English",
volume = "53",
pages = "75--84",
journal = "Annals of Clinical Biochemistry",
issn = "0004-5632",
publisher = "SAGE Publications Ltd",
number = "1",

}

TY - JOUR

T1 - Highly sensitive reversed-phase high-performance liquid chromatography assay for the detection of Tamm–Horsfall protein in human urine

AU - Akimoto, Masaru

AU - Hokazono, Eisaku

AU - Ota, Eri

AU - Tateishi, Takiko

AU - Kayamori, Yuzo

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Background: Tamm–Horsfall protein (also known as uromodulin) is the most abundant urinary protein in healthy individuals. Since initially characterized by Tamm and Horsfall, the amount of urinary excretion and structural mutations of Tamm–Horsfall protein is associated with kidney diseases. However, currently available assays for Tamm–Horsfall protein, which are mainly enzyme-linked immunosorbent assay-based, suffer from poor reproducibility and might give false negative results. Methods: We developed a novel, quantitative assay for Tamm–Horsfall protein using reversed-phase high-performance liquid chromatography. A precipitation pretreatment avoided urine matrix interference and excessive sample dilution. High-performance liquid chromatography optimization based on polarity allowed excellent separation of Tamm–Horsfall protein from other major urine components. Results: Our method exhibited high precision (based on the relative standard deviations of intraday [≤2.77%] and interday [≤5.35%] repetitions). The Tamm–Horsfall protein recovery rate was 100.0–104.2%. The mean Tamm–Horsfall protein concentration in 25 healthy individuals was 31.6 ± 18.8 mg/g creatinine. There was a strong correlation between data obtained by high-performance liquid chromatography and enzyme-linked immunosorbent assay (r = 0.906), but enzyme-linked immunosorbent assay values tended to be lower than high-performance liquid chromatography values at low Tamm–Horsfall protein concentrations. Conclusions: The high sensitivity and reproducibility of our Tamm–Horsfall protein assay will reduce the number of false negative results of the sample compared with enzyme-linked immunosorbent assay. Moreover, our method is superior to other high-performance liquid chromatography methods, and a simple protocol will facilitate further research on the physiological role of Tamm–Horsfall protein.

AB - Background: Tamm–Horsfall protein (also known as uromodulin) is the most abundant urinary protein in healthy individuals. Since initially characterized by Tamm and Horsfall, the amount of urinary excretion and structural mutations of Tamm–Horsfall protein is associated with kidney diseases. However, currently available assays for Tamm–Horsfall protein, which are mainly enzyme-linked immunosorbent assay-based, suffer from poor reproducibility and might give false negative results. Methods: We developed a novel, quantitative assay for Tamm–Horsfall protein using reversed-phase high-performance liquid chromatography. A precipitation pretreatment avoided urine matrix interference and excessive sample dilution. High-performance liquid chromatography optimization based on polarity allowed excellent separation of Tamm–Horsfall protein from other major urine components. Results: Our method exhibited high precision (based on the relative standard deviations of intraday [≤2.77%] and interday [≤5.35%] repetitions). The Tamm–Horsfall protein recovery rate was 100.0–104.2%. The mean Tamm–Horsfall protein concentration in 25 healthy individuals was 31.6 ± 18.8 mg/g creatinine. There was a strong correlation between data obtained by high-performance liquid chromatography and enzyme-linked immunosorbent assay (r = 0.906), but enzyme-linked immunosorbent assay values tended to be lower than high-performance liquid chromatography values at low Tamm–Horsfall protein concentrations. Conclusions: The high sensitivity and reproducibility of our Tamm–Horsfall protein assay will reduce the number of false negative results of the sample compared with enzyme-linked immunosorbent assay. Moreover, our method is superior to other high-performance liquid chromatography methods, and a simple protocol will facilitate further research on the physiological role of Tamm–Horsfall protein.

UR - http://www.scopus.com/inward/record.url?scp=84950336309&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84950336309&partnerID=8YFLogxK

U2 - 10.1177/0004563215583698

DO - 10.1177/0004563215583698

M3 - Article

C2 - 25838415

AN - SCOPUS:84950336309

VL - 53

SP - 75

EP - 84

JO - Annals of Clinical Biochemistry

JF - Annals of Clinical Biochemistry

SN - 0004-5632

IS - 1

ER -