Immunochemical property of human haptoglobin phenotypes: Determination of plasma haptoglobin using type-matched standards

Cheng, Tsai Mu; Pan, Ju Pin; Lai, Shiau Ting; Kao, Li Pin; Lin, Hong; Mao, Simon J.T.

doi:10.1016/j.clinbiochem.2007.04.018

Cited by 28 publications

(55 citation statements)

References 38 publications

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“…Then, the mixture was placed on a 7% native polyacrylamide gel (pH 8.8), and electrophoresis was conducted with a voltage range of 120–150 V. After electrophoresis completion, the Hp-Hb complex was visualized by shaking the gel in freshly prepared peroxidase substrate with 0.05% (w/v) 3,3’- diaminobenzidine and 0.07% (v/v) hydrogen peroxide. Hp phenotyping results were confirmed by Western blotting using an α-chain specific monoclonal antibody [15]. …”

Section: Methodsmentioning

confidence: 99%

“…Hp phenotyping was performed using a native polyacrylamide gel electrophoresis with Hb-supplemented plasma [15]. A certain amount of plasma sample (7 μL) was mixed with 8 mg/mL Hb (5 μL) and equilibrated with sample buffer (3 μL) containing 0.625 mol/L Tris-base, 0.125 mg/L bromophenol blue and 50% (v/v) glycerol.…”

Section: Methodsmentioning

confidence: 99%

“…Plasma levels of Hp were measured using a Human Hp Quantikine ELISA Kit based on the manufacturer’s instructions (R&D Systems Inc., Minneapolis, MN, USA) [17]. …”

Section: Methodsmentioning

confidence: 99%

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Elevated Plasma Haptoglobin Level as a Potential Marker for Poor Prognosis in Acute Cerebral Infarction

Zhang

Xiang

2018

Eur Neurol

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Background: Relationship between peripheral circulation concentration of haptoglobin (Hp) and risk of cerebrovascular disease has not been well investigated so far. The aim of the study was to reveal the prognostic significance of Hp on acute cerebral infarction. Methods: A total of 200 patients with acute cerebral infarction and 200 controls were included. Research data were collected from their medical records. Blood samples were obtained on admission. Hp phenotyping was performed using a native polyacrylamide gel electrophoresis. Plasma Hp levels were determined using enzyme-linked immunosorbent assays. The patients were followed up yearly, and endpoint was overall death. Results: Plasma levels of Hp were significantly elevated in the patients than in the controls (p < 0.001). Three-year overall death in the follow-up period was associated with the high plasma level of Hp (Hazard ratio [HR] 2.33, 95% CI 1.74–3.12). But, 3-year overall death was not related to Hp phenotyping (Hp 2–1 vs. 1–1: HR 1.07, 95% CI 0.66–1.73; Hp 2–2 vs. 1–1: HR 1.08, 95% CI 0.67–1.74). Conclusion: Peripheral circulation concentration of Hp might be an independent prognostic factor for acute cerebral infarction. But there was no relationship between Hp phenotyping and prognosis in this disease.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Elevated Plasma Haptoglobin Level as a Potential Marker for Poor Prognosis in Acute Cerebral Infarction

Zhang

Xiang

2018

Eur Neurol

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“…We have recently developed several lines of human Hp mAb and routinely utilized these antibodies for the isolation of human Hp 1-1, 2-1 and 2-2 phenotypes [22,26]. As only W1 (specific to the a-chain) is able to cross-react with the deer a-chain on a western blot, we attempted to utilize this mAb for the affinity isolation of deer Hp in this study.…”

Section: Isolation Of Deer Native Hpmentioning

confidence: 99%

A unique tetrameric structure of deer plasma haptoglobin – an evolutionary advantage in the Hp 2‐2 phenotype with homogeneous structure

et al. 2008

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Haptoglobin (Hp) is an acute-phase protein (responsive to infection and inflammation) that is present in the plasma of all mammals [1][2][3][4]. A recent study has found that Hp also exists in lower vertebrates (bony fish) but not in frog and chicken [5]. The most frequently reported biological functions of the protein are to capture released hemoglobin during excessive hemolysis [6] and to scavenge free radicals during oxidative Similar to blood types, human plasma haptoglobin (Hp) is classified into three phenotypes: Hp 1-1, 2-1 and 2-2. They are genetically inherited from two alleles Hp 1 and Hp 2 (represented in bold), but only the Hp 1-1 phenotype is found in almost all animal species. The Hp 2-2 protein consists of complicated large polymers cross-linked by a2-b subunits or (a2-b) n (where n ‡ 3, up to 12 or more), and is associated with the risk of the development of diabetic, cardiovascular and inflammatory diseases. In the present study, we found that deer plasma Hp mimics human Hp 2, containing a tandem repeat over the a-chain based on our cloned cDNA sequence. Interestingly, the isolated deer Hp is homogeneous and tetrameric, i.e. (a-b) 4 , although the locations of )SH groups (responsible for the formation of polymers) are exactly identical to that of human. Denaturation of deer Hp using 6 m urea under reducing conditions (143 mm b-mercaptoethanol), followed by renaturation, sustained the formation of (a-b) 4 , suggesting that the Hp tetramers are not randomly assembled. Interestingly, an a-chain monoclonal antibody (W1), known to recognize both human and deer a-chains, only binds to intact human Hp polymers, but not to deer Hp tetramers. This implies that the epitope of the deer a-chain is no longer exposed on the surface when Hp tetramers are formed. We propose that steric hindrance plays a major role in determining the polymeric formation in human and deer polymers. Phylogenetic and immunochemical analyses revealed that the Hp 2 allele of deer might have arisen at least 25 million years ago. A mechanism involved in forming Hp tetramers is proposed and discussed, and the possibility is raised that the evolved tetrameric structure of deer Hp might confer a physiological advantage.Abbreviations Hp, haptoglobin; b-ME, b-mercaptoethanol.

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“…In ELISA, the activity of the enzyme attached to anti-Hp antibody can consequently be measured by means of its reaction with a chromogenic substrate to generate a measurable signal, which can be correlated to the amount of Hp present in a sample. Other than ELISA, electrochemical impedance spectroscopy, 3 time-resolved immune fluorometry, 4 labor-intensive electrophoresis, chromogen staining, 5 or chemiluminescent imaging 6 have also been reported for Hp quantification. All these methods are very much time consuming and labor intensive for clinical applications.…”

Section: Introductionmentioning

confidence: 99%

SERS-based quantitative detection of ovarian cancer prognostic factor haptoglobin

Perumal¹,

Balasundaram²,

Mahyuddin³

et al. 2015

IJN

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Surface-enhanced Raman spectroscopy (SERS) is increasingly being used for biosensing because of its high sensitivity and low detection limit, which are made possible by the unique Raman ‘fingerprint’ spectra from the biomolecules. Here we propose a novel SERS method for the fast, sensitive, and reliable quantitative analysis of haptoglobin (Hp), an acute phase plasma glycoprotein that is widely gaining application as a prognostic ovarian cancer biomarker. We exploited the peroxidase activity of the hemoglobin–haptoglobin (Hb–Hp) complex formed by the selective and specific binding of Hp to free Hb to catalyze the reaction of 3,3′,5,5′-tetramethylbenzidine (TMB) substrate and hydrogen peroxide to result in the final product of strongly SERS-active TMB 2+ . We observed a linear increase in the SERS signal of TMB 2+ with increasing concentrations of Hb–Hp complex from 50 nM to 34 μM. Based on this concentration-dependent SERS spectrum, we quantified Hp in clinical samples. We observed that our inference about the prognosis of the disease coincided with the histology data and that our method was much more sensitive than the enzyme-linked immunosorbent assay method.

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Immunochemical property of human haptoglobin phenotypes: Determination of plasma haptoglobin using type-matched standards

Cited by 28 publications

References 38 publications

Elevated Plasma Haptoglobin Level as a Potential Marker for Poor Prognosis in Acute Cerebral Infarction

Elevated Plasma Haptoglobin Level as a Potential Marker for Poor Prognosis in Acute Cerebral Infarction

A unique tetrameric structure of deer plasma haptoglobin – an evolutionary advantage in the Hp 2‐2 phenotype with homogeneous structure

SERS-based quantitative detection of ovarian cancer prognostic factor haptoglobin

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