Rapid quantitation of human epididymis protein 4 in human serum by amplified luminescent proximity homogeneous immunoassay (AlphaLISA)

Zhao, Hui; Lin, Guanfeng; Liu, Tiancai; Liang, Junyu; Ren, Zhi-Qi; Liang, Rongliang; Chen, Baihong; Huang, Wenhua; Wu, Yixuan

doi:10.1016/j.jim.2016.08.006

Cited by 25 publications

(13 citation statements)

References 26 publications

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“…AlphaLISA technology is a bead-based method relying on the interaction between donor beads and acceptor beads. When the antibody-antigen reaction causes donor beads and acceptor beads to approach each other, the laser excites the cascade reaction, resulting in a greatly amplified signal [ 19 , 20 ]. AlphaLISA method significantly improves the sensitivity of detection with obvious advantages and can be used as a powerful assay method for the development of ENO1 detection in the peripheral blood.…”

Section: Discussionmentioning

confidence: 99%

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ENO1 Overexpression in Pancreatic Cancer Patients and Its Clinical and Diagnostic Significance

Yin

Wang

Liu

2018

Gastroenterology Research and Practice

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We investigated in this study the expression of ENO1 in tissues and plasma of PDAC patients to evaluate its clinicopathological and diagnostic significance. ENO1 protein expression was detected in tissue microarray of human PDAC and adjacent noncancer tissues. Electrochemiluminescence immunoassay and amplified luminescent proximity homogeneous assay (AlphaLISA) were performed to measure CA19-9 and ENO1 concentration in plasma from PDAC patients and healthy controls. We demonstrated that ENO1 overexpression is positively correlated with clinical stage, lymph node metastasis, and poor prognosis of PDAC; ENO1 may function as a hopeful candidate diagnostic marker in combination with CA19-9 in PDAC diagnosis.

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Section: Discussionmentioning

confidence: 99%

“…Plasma concentration of ENO1 was determined by amplified luminescent proximity homogeneous assay (AlphaLISA) method as described previously [ 19 , 20 ]. Anti-ENO1 monoclonal antibodies (WH0002023M1, SAB1403772) and polyclonal antibodies (AV34376, E2659) were obtained from Sigma-Aldrich (St. Louis, MO, USA).…”

Section: Methodsmentioning

confidence: 99%

ENO1 Overexpression in Pancreatic Cancer Patients and Its Clinical and Diagnostic Significance

Yin

Wang

Liu

2018

Gastroenterology Research and Practice

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“…Therefore, these components might be potential pitfalls of assays based in luminescence response. However, it has already been demonstrated by several authors that AlphaLISA ® assays are unaffected by interfering with antioxidant components as ascorbic acid or bilirubin as well as triglycerides contained in plasma samples (29, 35). In addition, here we have also tested that prooxidant components as H 2 O 2 did not alter the level of RLU observed for the MMP-9–TIMP-1 interactions using rMMP-9 and TIMP-1 proteins (Figure S1A in Supplementary Material) or endogenous MMP-9–TIMP-1 interactions in human plasma samples (Figure S1B in Supplementary Material).…”

Section: Anticipated Resultsmentioning

confidence: 99%

Rapid, Automated, and Specific Immunoassay to Directly Measure Matrix Metalloproteinase-9–Tissue Inhibitor of Metalloproteinase-1 Interactions in Human Plasma Using AlphaLISA Technology: A New Alternative to Classical ELISA

et al. 2017

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The protocol describes a novel, rapid, and no-wash one-step immunoassay for highly sensitive and direct detection of the complexes between matrix metalloproteinases (MMPs) and their tissue inhibitor of metalloproteinases (TIMPs) based on AlphaLISA® technology. We describe two procedures: (i) one approach is used to analyze MMP-9–TIMP-1 interactions using recombinant human MMP-9 with its corresponding recombinant human TIMP-1 inhibitor and (ii) the second approach is used to analyze native or endogenous MMP-9–TIMP-1 protein interactions in samples of human plasma. Evaluating native MMP-9–TIMP-1 complexes using this approach avoids the use of indirect calculations of the MMP-9/TIMP-1 ratio for which independent MMP-9 and TIMP-1 quantifications by two conventional ELISAs are needed. The MMP-9–TIMP-1 AlphaLISA® assay is quick, highly simplified, and cost-effective and can be completed in less than 3 h. Moreover, the assay has great potential for use in basic and preclinical research as it allows direct determination of native MMP-9–TIMP-1 complexes in circulating blood as biofluid.

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“…Recently, human epididymis protein-4, also named as whey-acidic-protein four-disulfide core protein-2 (WFDC2), is one of the most promising new biomarkers [9] , [10] and has been approved by the Food and Drug Administration (FDA) as the sensitive serum biomarkers for the early diagnosis and monitoring of epithelial ovarian cancer. To date, various analysis methods have been widely established for the serum HE4 detection, including enzyme-linked immunosorbent assay (ELISA), electrochemiluminescence enzyme immunoassay, time-resolved fluoroimmunoassay (TRFIA), amplified luminescent proximity homogeneous immunoassay (AlphaLISA) [11] , [12] , [13] , [14] . Despite the considerable advancements in technology, many disadvantages still exist.…”

Section: Introductionmentioning

confidence: 99%

The fabrication of magnetic particle-based chemiluminescence immunoassay for human epididymis protein-4 detection in ovarian cancer

Liu

Qiu

et al. 2018

Biochemistry and Biophysics Reports

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The magnetic particles have a significant influence on the immunoassay detection and cancer therapy. Herein, the chemiluminescence immunoassay combined with the magnetic particles (MPCLIA) was presented for the clinical determination and analysis of human epididymis protein 4 (HE4) in the human serum. Under the optimized experiment conditions, the secure MPCLIA method can detect HE4 in the broader range of 0–1000 pmol/L, with a lower detection limit of 1.35 pmol/L. The satisfactory recovery rate of the method in the serum ranged from 83.62% to 105.10%, which was well within the requirement of clinical analysis. Moreover, the results showed the good correlation with enzyme-linked immunosorbent assay (ELISA), with the correlation coefficient of 0.9589. This proposed method has been successfully applied to the clinical determination of HE4 in the human serum.

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Rapid quantitation of human epididymis protein 4 in human serum by amplified luminescent proximity homogeneous immunoassay (AlphaLISA)

Cited by 25 publications

References 26 publications

ENO1 Overexpression in Pancreatic Cancer Patients and Its Clinical and Diagnostic Significance

ENO1 Overexpression in Pancreatic Cancer Patients and Its Clinical and Diagnostic Significance

Rapid, Automated, and Specific Immunoassay to Directly Measure Matrix Metalloproteinase-9–Tissue Inhibitor of Metalloproteinase-1 Interactions in Human Plasma Using AlphaLISA Technology: A New Alternative to Classical ELISA

The fabrication of magnetic particle-based chemiluminescence immunoassay for human epididymis protein-4 detection in ovarian cancer

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