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Micromachines 2017, 8(5), 149; doi:10.3390/mi8050149

Preparation of an Acridinium Ester-Labeled Antibody and Its Application in GoldMag Nanoparticle-Based, Ultrasensitive Chemiluminescence Immunoassay for the Detection of Human Epididymis Protein 4

1
,
2
,
3
,
1,2,* and 1,2,*
1
College of Life Sciences, Northwest University, Xi’an 710069, China
2
National Engineering Research Center for Miniaturized Detection Systems, Xi’an 710069, China
3
Department of Biology, Providence College, Providence, RI 02912, USA
*
Authors to whom correspondence should be addressed.
Academic Editors: Roger Jagdish Narayan, Adrian Martinez-Rivas and Marco Ramírez Salinas
Received: 20 March 2017 / Revised: 26 April 2017 / Accepted: 28 April 2017 / Published: 7 May 2017
(This article belongs to the Special Issue Medical Microdevices and Micromachines)
View Full-Text   |   Download PDF [2950 KB, uploaded 7 May 2017]   |  

Abstract

An ultrasensitive and rapid sandwich-type chemiluminescence immunoassay (CLIA) was developed for the clinical determination of human epididymis protein 4 (HE4) in human serum, using GoldMag nanoparticles as solid phase and acridinium ester (AE) as chemiluminescence system (GMP-CLIA). The process of AE labeling antibodies was systematically studied and evaluated. The effect of varies factors such as molar ratio of AE to antibodies, labeling time, and the components of elution buffer and trigger solution were optimized. Under the selected conditions, AE labeling experiments were successfully performed with the average labeling efficiency of 1.92 ± 0.08, and antibody utilization rate of 69.77 ± 1.19%. Antibody activity remained unchanged after labeling. The established GMP-CLIA method can detect HE4 in the range of 0.25–50 ng·mL−1 (10–2000 pM) with a detection limit of 0.084 ng·mL−1 (3.36 pM). The sensitivity has reached a high level, comparable with the current commercial detection kits. This proposed method has been successfully applied to the clinical determination of HE4 in 65 human sera. The results showed a good correlation with a clinical method, microplate-based chemiluminescence enzyme immunoassay (CLEIA), with the correlation coefficient of 0.9594. View Full-Text
Keywords: GoldMag particles; chemiluminescence immunoassay; acridinium ester; labeling; human epididymis protein 4 GoldMag particles; chemiluminescence immunoassay; acridinium ester; labeling; human epididymis protein 4
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Ma, T.; Zhang, M.; Wan, Y.; Cui, Y.; Ma, L. Preparation of an Acridinium Ester-Labeled Antibody and Its Application in GoldMag Nanoparticle-Based, Ultrasensitive Chemiluminescence Immunoassay for the Detection of Human Epididymis Protein 4. Micromachines 2017, 8, 149.

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