Fluorescent protein-based detection of unconjugated bilirubin in newborn serum

Iwatani, Sota; Nakamura, Hajime; Kurokawa, Daisuke; Yamana, Kazunari; Nishida, Kosuke; Fukushima, Sachiyo; Koda, Tsubasa; Nishimura, Noriyuki; Nishio, Hisahide; Iijima, Kazumoto; Miyawaki, Atsushi; Morioka, Ichiro

doi:10.1038/srep28489

Cited by 28 publications

(37 citation statements)

References 23 publications

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“…The nanosheets with different layer influence the accuracy of the linear law. Most of the previously reported BR fluorescent probes or sensors are based on fluorescent proteins [39], metal organic frameworks (MOFs) [40], quantum dots [38], and rare-earth complexes [42]. The use of fluorescent proteins offers the advantages of a broad detection range and good linearity, but imposes strict preparation and storage requirements and incurs higher costs than other methods.…”

Section: Response Of the Sensing Film To Brmentioning

confidence: 99%

Luminescent sensing film based on sulfosalicylic acid modified Tb(III)-doped yttrium hydroxide nanosheets

Yang

Zheng

et al. 2018

J Adv Ceram

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Sulfosalicylic acid (SSA) was used as an intercalation agent and an excellent antenna to synthesize layered rare-earth hydroxide (LRH) materials and directly obtain SSA-modified terbium-doped ytterbium hydroxide nanosheets by mechanical exfoliation. The crystal structure and morphologies of the LRHs and nanosheets were determined by X-ray diffraction, scanning electron microscopy, and transmission electron microscopy. The particle size and zeta potential of the prepared nanosheets were also analyzed. The as-prepared nanosheets exhibited excellent luminescent properties. The positively charged nanosheets were electrophoretically deposited on a conductive glass to form a thin film. The luminescence of this thin film can be quenched by chromate (CrO 4 2-) and bilirubin (BR), which shows good sensing properties. The quenching mechanism of the sensing film by CrO 4 2and BR was discussed based on the spectra and structure of the film.

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Section: Response Of the Sensing Film To Brmentioning

confidence: 99%

Luminescent sensing film based on sulfosalicylic acid modified Tb(III)-doped yttrium hydroxide nanosheets

Yang

Zheng

et al. 2018

J Adv Ceram

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“…Other methods for bilirubin determination comprise high performance liquid chromatography (HPLC)‐based procedures, enzymatic reactions and spectrophotometric analyses (Puppalwar et al, ). However, assays that enable the direct measure of bilirubin are considered superior to current tests (Iwatani et al, ; Kumagai et al, ).…”

Section: Resultsmentioning

confidence: 99%

“…However, assays that enable the direct measure of bilirubin are considered superior to current tests (Iwatani et al, 2016;Kumagai et al, 2013).…”

Section: Functional Characterization Of the Hug Proteinmentioning

confidence: 99%

Human elastin‐like polypeptides as a versatile platform for exploitation of ultrasensitive bilirubin detection by UnaG

Bandiera

Corich

Tommasi

et al. 2019

Biotech & Bioengineering

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A new, bifunctional recombinant protein was expressed as the fusion product of human elastin‐like polypeptide (HELP) and the bilirubin‐binding protein UnaG. The engineered product displays both the HELP‐specific property of forming a functional hydrogel matrix and the UnaG‐specific capacity of emitting green fluorescence upon ligand binding. The new fusion protein has been proven to be effective at detecting bilirubin in complex environments with high background noise. A cell culture model of the stress response, consisting of bilirubin released in the cell culture medium, was set up to assess the bilirubin‐sensing properties of the functional matrix obtained by cross‐linking the HELP moiety. Our engineered protein allowed us to monitor cell induction by the release of bilirubin in the culture medium on a nanomolar scale. This study shows that elastin‐like protein fusion represents a versatile platform for the development of novel and commercially viable analytical and biosensing devices.

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“…UB is calculated from the initial velocity of bilirubin degradation and the ratio of the POD concentration to that in the standard assay of the albumin-free bilirubin solution [ 1 , 6 , 7 ] ( Figure 5 a). As UnaG is a protein characterized by iDB concentration-dependent fluorescence [ 9 , 10 ], the GOD–POD–UnaG method involves the division of the serum sample into two samples to calculate the difference in TB level initially and after 20 s of POD reaction (TB reduction rate). Therefore, it is necessary to stop the POD reaction after 20 s, which can be accomplished by adding ascorbic acid (patent registration number: 6716108) ( Figure 5 b).…”

Section: Discussionmentioning

confidence: 99%

A Novel Method for Measuring Serum Unbound Bilirubin Levels Using Glucose Oxidase–Peroxidase and Bilirubin-Inducible Fluorescent Protein (UnaG): No Influence of Direct Bilirubin

Iwatani

Yamana

Nakamura

et al. 2020

IJMS

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The glucose oxidase–peroxidase (GOD–POD) method used to measure serum unbound bilirubin (UB) suffers from direct bilirubin (DB) interference. Using a bilirubin-inducible fluorescent protein from eel muscle (UnaG), a novel GOD–POD–UnaG method for measuring UB was developed. Newborn sera with an indirect bilirubin/albumin (iDB/A) molar ratio of <0.5 were classified into four groups of DB/total serum bilirubin (TB) ratios (<5%, 5–10%, 10–20%, and ≥20%), and the correlation between the UB levels and iDB/A ratio was examined. Linear regression analysis was performed to compare UB values from both methods with the iDB/A ratio from 38 sera samples with DB/TB ratio <5% and 11 samples with DB/TB ratio ≥5%. The correlation coefficient (r) between UB values and the iDB/A ratio for the GOD–POD method was 0.8096 (DB/TB ratio <5%, n = 239), 0.7265 (5–10%, n = 29), 0.7165 (10–20%, n = 17), and 0.4816 (≥20%, n = 16). UB values using the GOD–POD–UnaG method highly correlated with the iDB/A ratio in both <5% and ≥5% DB/TB ratio sera (r = 0.887 and 0.806, respectively), whereas a low correlation (r = 0.428) occurred for ≥5% DB/TB ratio sera using the GOD–POD method. Our GOD–POD–UnaG method can measure UB levels regardless of the presence of DB.

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Fluorescent protein-based detection of unconjugated bilirubin in newborn serum

Cited by 28 publications

References 23 publications

Luminescent sensing film based on sulfosalicylic acid modified Tb(III)-doped yttrium hydroxide nanosheets

Luminescent sensing film based on sulfosalicylic acid modified Tb(III)-doped yttrium hydroxide nanosheets

Human elastin‐like polypeptides as a versatile platform for exploitation of ultrasensitive bilirubin detection by UnaG

A Novel Method for Measuring Serum Unbound Bilirubin Levels Using Glucose Oxidase–Peroxidase and Bilirubin-Inducible Fluorescent Protein (UnaG): No Influence of Direct Bilirubin

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