Liquid-Phase Binding Assay of α-Fetoprotein Using a Sulfated Antibody for Bound/Free Separation

Nakamura, Kenji; Imajo, Nobuko; Yamagata, Yukari; Katoh, Hideo; Fujio, K.; Tanaka, Takumi; Satomura, Shinji; Matsuura, Shuji

doi:10.1021/ac9711495

Cited by 21 publications

(14 citation statements)

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“…Serum des-gamma-carboxy prothrombin (DCP) was measured using a l-TAS assay for DCP (Wako Pure Chemical Industries Ltd, Osaka, Japan). The lower limits of quantitation for l-TAS assay AFP (L1 and L3), LiBASys assay AFP (L1 and L3), and l-TAS assay DCP are 0.3 ng/ml AFP (L1 and L3), 0.8 ng/ml AFP (L1 and L3), and 5 mAU/ml DCP, respectively [14][15][16]. When AFP-L3 is not detectable, the percentage of AFP-L3 is defined as 0.5%.…”

Section: Laboratory Examinationmentioning

confidence: 96%

“…As a reference method, AFP-L1 and AFP-L3 were also measured using a liquid-phase binding assay system (LiBASys) at the same time (Wako Pure Chemical Industries Ltd., Osaka, Japan) [14]. Total AFP concentration (ng/ ml) in the serum sample was determined by addition of AFP-L3 to AFP-L1.…”

Section: Laboratory Examinationmentioning

confidence: 99%

“…Moreover, its level predicts the malignant potential of HCC with subsequent unfavorable prognosis after treatment [9][10][11][12][13]. However, measurement of AFP-L3 has not always been reliable for serum samples with low total AFP concentration determined by conventional lectin affinity electrophoresis or using a liquid-phase binding assay system (LiBASys) [14].…”

Section: Introductionmentioning

confidence: 99%

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Clinical Advantage of Highly Sensitive On-Chip Immunoassay for Fucosylated Fraction of Alpha-Fetoprotein in Patients with Hepatocellular Carcinoma

et al. 2010

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Section: Laboratory Examinationmentioning

confidence: 96%

Section: Laboratory Examinationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Clinical Advantage of Highly Sensitive On-Chip Immunoassay for Fucosylated Fraction of Alpha-Fetoprotein in Patients with Hepatocellular Carcinoma

et al. 2010

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“…Highly sensitive AFP-L3% (hs-AFP-L3%) were measured by a microchip capillary electrophoresis and liquid-phase binding assay on a µ-TASWako i30 auto analyzer (Wako Pure Chemical Industries, Ltd., Osaka, Japan) (15). Conventional AFP-L3% (c-AFP-L3%) was examined using a column chromatography and liquidphase binding assay on a LiBASys auto analyzer (Wako Pure Chemical Industries, Ltd.) (13). The analytical sensitivity of the µ-TASWako i30 auto analyzer is 0.3 µg/ml AFP; the AFP-L3% can be measured when AFP-L3 is over 0.3 µg/ml.…”

Section: Measurement Of Serum Afp and Afp-l3%mentioning

confidence: 99%

“…Therefore, it is not clear how useful this test is for the early detection of HCC. Additionally, measurement of AFP-L3 has not always been reliable for serum samples with low total AFP concentration, as determined by conventional lectin affinity system (LiBASys) (13).…”

Section: Introductionmentioning

confidence: 99%

Highly sensitive lens culinaris agglutinin-reactive α-fetoprotein is useful for early detection of hepatocellular carcinoma in patients with chronic liver disease

Ido

2011

Oncol Rep

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Abstract. The fucosylated fraction of α-fetoprotein (AFP-L3) is a specific marker for hepatocellular carcinoma (HCC). However, conventional AFP-L3% (c-AFP-L3%) has not always been reliable in cases with low serum α-fetoprotein (AFP) levels. In this study, we evaluated the clinical utility of a newly developed assay, highly sensitive AFP-L3% (hs-AFP-L3%). Subjects included 74 patients with benign liver disease (BLD), including chronic hepatitis and cirrhosis, and 94 with HCC. Serum hs-AFP-L3% was significantly higher than c-AFP-L3% in patients with early-stage HCC (solitary or <20 mm in diameter). Additionally, hs-AFP-L3% was significantly increased in patients with well-differentiated HCC. In patients with serum AFP <20 ng/ml, the sensitivities of c-AFP-L3% and hs-AFP-L3% were 12.5 and 44.6%, respectively, at a cut-off value of 5%. In 59 BLD patients with serum AFP <20 ng/ml, the HCC-positive rate in patients with hs-AFP-L3% ≥5% was significantly higher compared to those with hs-AFP-L3% <5% during the follow-up period (median, 35 months; range, 5-48 months). Importantly, none of the BLD patients with both serum AFP <20 ng/ml and hs-AFP-L3% <5% developed HCC. These results indicated that hs-AFP-L3% is useful for early detection of HCC in BLD patients, even for those with serum AFP <20 ng/ml. Furthermore, since hs-AFP-L3% increases before HCC is detectable by various advanced imaging modalities, this assay may help identify BLD patients with a higher risk of HCC. IntroductionHepatocellular carcinoma (HCC) is the sixth most common cancer in the world, and the third most common cause of cancer-related death (1). Although it is more common in Asia and Africa, its incidence in the United States has increased over the past two decades, largely due to the spread of hepatitis C (HCV) infection, which is an underlying risk factor (2). Early detection of HCC increases the potential for curative treatment and improves prognosis. Several methods developed for the diagnosis of HCC, including evaluation of serum markers, ultrasonography (US), computed tomography (CT) and magnetic resonance imaging (MRI), have been tested clinically. α-fetoprotein (AFP) and des-γ carboxy prothrombin (DCP), serum proteins that are elevated in HCC, are the most widely used markers. Although routine screening offers the best chance for early tumor detection, the reported sensitivities and specificities of elevated serum AFP and DCP levels vary significantly (3-8). Furthermore, serum AFP levels increase in only 30-40% of patients with HCC, especially early in the disease process (5). Additionally, an increase in serum AFP is also seen in patients with non-cancerous conditions, including cirrhosis or exacerbation of chronic hepatitis (9). AFP-L3, the lectin lens culinaris agglutinin-bound fraction, is one of the three glycoforms of AFP, and is the major glycoform elevated in the serum of HCC patients. The reported sensitivities of AFP-L3 as a method of detecting HCC range from 75-97% with specificities of 90-92% (10,11). In cases of HCC, however, h...

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Development of on‐chip fully automated immunoassay system “μTASWako i30” to measure the changes in glycosylation profiles of alpha‐fetoprotein in patients with hepatocellular carcinoma

Kurosawa¹,

Watanabe²

2016

Proteomics

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Glycosylation profiles significantly change during oncogenesis. Aberrant glycosylation can be used as a cancer biomarker in clinical settings. Different glycoforms can be separately detected using lectin affinity electrophoresis and lectin array-based methods. However, most methodologies and procedures need experienced technique to perform the assays and expertise to interpret the results. To apply glycomarkers for clinical practice, a robust assay system with an easy-to-use workflow is required. Wako's μTASWako i30, a fully automated immunoanalyzer, was developed for in vitro diagnostics based on microfluidic technology. It utilizes the principles of liquid-phase binding assay, where immunoreactions are performed in a liquid phase, and electrokinetic analyte transport assay. Capillary electrophoresis on microfluidic chip has enabled the detection of different glycoform types of alpha-fetoprotein (AFP), a serum biomarker for hepatocellular carcinoma. AFP with altered glycosylation can be separated based on the reactivity to Lens culinaris agglutinin on electrophoresis. The glycoform AFP-L3 was reportedly more specific in hepatocellular carcinoma. This assay system can provide a high sensitivity and rapid results in 9 min. The test results for ratio of AFP-L3 to total AFP using μTASWako i30 are correlated with those of conventional methodology. The μTASWako assay system and the technology can be utilized for glycosylation analysis in the postgenomic era.

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Liquid-Phase Binding Assay of α-Fetoprotein Using a Sulfated Antibody for Bound/Free Separation

Cited by 21 publications

References 5 publications

Clinical Advantage of Highly Sensitive On-Chip Immunoassay for Fucosylated Fraction of Alpha-Fetoprotein in Patients with Hepatocellular Carcinoma

Clinical Advantage of Highly Sensitive On-Chip Immunoassay for Fucosylated Fraction of Alpha-Fetoprotein in Patients with Hepatocellular Carcinoma

Highly sensitive lens culinaris agglutinin-reactive α-fetoprotein is useful for early detection of hepatocellular carcinoma in patients with chronic liver disease

Development of on‐chip fully automated immunoassay system “μTASWako i30” to measure the changes in glycosylation profiles of alpha‐fetoprotein in patients with hepatocellular carcinoma

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