Enzymatic assay for conjugated bilirubin (Bc) in serum using bilirubin oxidase (BOD)

Kimura, Shigeki; Iyama, Shigeru; Yamaguchi, Yoshihiro; Hayashi, S.; Yanagihara, Takehiko

doi:10.1002/(sici)1098-2825(1999)13:5<219::aid-jcla5>3.0.co;2-6

Cited by 8 publications

(7 citation statements)

References 16 publications

(18 reference statements)

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“…This is the first large-scale clinical study demonstrating the applicability of eel-derived fluorescent protein for measurement of unconjugated bilirubin concentrations in newborn serum samples. The concentrations measured by this method showed high correlation with those obtained by the conventional bilirubin oxidase method, which has itself shown good correlation with those obtained by high-performance liquid chromatography, considered as the gold standard 12 14 15 . The performance of the UnaG method was unaffected by PT and the presence of serum hemoglobin and lipid emulsion.…”

Section: Discussionsupporting

confidence: 56%

“…The concentrations measured by this method showed high correlation with those obtained by the conventional bilirubin oxidase method, which has itself shown good correlation with those obtained by high-performance liquid chromatography, considered as the gold standard121415. The performance of the UnaG method was unaffected by PT and the presence of serum hemoglobin and lipid emulsion.…”

Section: Discussionsupporting

confidence: 52%

“…Total and conjugated bilirubin concentrations in serum were measured using IatroLQ T-bil and IatroLQ D-bil kits (Unitika Co., Okazaki, Japan) as previously reported1415. Serum unconjugated bilirubin concentration was calculated with the following formula (1 mg/dl = 17.1 μM): [unconjugated bilirubin] = [TB] – [conjugated bilirubin].…”

Section: Methodsmentioning

confidence: 99%

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Fluorescent protein-based detection of unconjugated bilirubin in newborn serum

Iwatani

Nakamura

Kurokawa

et al. 2016

Sci Rep

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Increased serum levels of unconjugated bilirubin are associated with the development of brain damage in newborns. In current clinical settings, there are no methods for directly determining serum levels of unconjugated bilirubin. UnaG, a fluorescent protein from Japanese eel muscle that specifically binds to unconjugated bilirubin was used in this study. Linear regression analysis was carried out to compare unconjugated bilirubin levels measured by UnaG and conventional bilirubin oxidase methods. Unconjugated bilirubin levels in the serum of newborns who were untreated or treated with phototherapy were compared. Effects of interfering factors in the serum (conjugated bilirubin, hemoglobin, and lipid) on unconjugated bilirubin concentration measured by the UnaG method were also evaluated. Unconjugated bilirubin levels measured by the UnaG method were highly correlated with those determined by the bilirubin oxidase assay. Unconjugated bilirubin levels determined by bilirubin oxidase and UnaG assays were similar in serum samples containing conjugated bilirubin. The performance of the UnaG assay was unaffected by phototherapy and the presence of serum hemoglobin and lipid emulsion. These results demonstrate the clinical applicability of the UnaG method for direct measurement of unconjugated bilirubin levels in newborn serum.

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Section: Discussionsupporting

confidence: 56%

Section: Discussionsupporting

confidence: 52%

Section: Methodsmentioning

confidence: 99%

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Fluorescent protein-based detection of unconjugated bilirubin in newborn serum

Iwatani

Nakamura

Kurokawa

et al. 2016

Sci Rep

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“…Serum UB was measured at a single peroxidase concentration as recommended by the manufacturer. Serum Alb and DB concentrations were measured using the modified bromocresol purple method and the bilirubin oxidase method, respectively 14,15 …”

Section: Methodsmentioning

confidence: 99%

Is bilirubin/albumin ratio correlated with unbound bilirubin concentration?

Sato

Morioka

Miwa³

et al. 2011

Pediatrics International

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“…Mv BOx can be utilized for a range of purposes. In medicine, it is used for diagnostics of bilirubin in serum 19–21 , in biotechnology for decolorization of synthetic dyes or detoxification of the environment 19,22,23 . It also shows a potential for use in biosensors and biofuel technology 24–33 .…”

Section: Introductionmentioning

confidence: 99%

Trp–His covalent adduct in bilirubin oxidase is crucial for effective bilirubin binding but has a minor role in electron transfer

Kovaĺ

Švecová

Østergaard

et al. 2019

Sci Rep

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Unlike any protein studied so far, the active site of bilirubin oxidase from Myrothecium verrucaria contains a unique type of covalent link between tryptophan and histidine side chains. The role of this post-translational modification in substrate binding and oxidation is not sufficiently understood. Our structural and mutational studies provide evidence that this Trp396–His398 adduct modifies T1 copper coordination and is an important part of the substrate binding and oxidation site. The presence of the adduct is crucial for oxidation of substituted phenols and it substantially influences the rate of oxidation of bilirubin. Additionally, we bring the first structure of bilirubin oxidase in complex with one of its products, ferricyanide ion, interacting with the modified tryptophan side chain, Arg356 and the active site-forming loop 393-398. The results imply that structurally and chemically distinct types of substrates, including bilirubin, utilize the Trp–His adduct mainly for binding and to a smaller extent for electron transfer.

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Enzymatic assay for conjugated bilirubin (Bc) in serum using bilirubin oxidase (BOD)

Cited by 8 publications

References 16 publications

Fluorescent protein-based detection of unconjugated bilirubin in newborn serum

Fluorescent protein-based detection of unconjugated bilirubin in newborn serum

Is bilirubin/albumin ratio correlated with unbound bilirubin concentration?

Trp–His covalent adduct in bilirubin oxidase is crucial for effective bilirubin binding but has a minor role in electron transfer

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