Highly Sensitive Fluorescence-Linked Immunosorbent Assay for the Determination of Human IgG in Serum Using Quantum Dot Nanobeads and Magnetic Fe(3)O(4) Nanospheres

Guo, Jiaping; Wang, Yameng; Niu, Shanshan; Li, Hongping; Tian, Yongmei; Yu, Songcheng; Yu, Fei; Wu, Yongjun; Liu, Li-e

[Image: see text] The aim of this study is to establish a new method with high sensitivity, accuracy, and stability for the determination of human IgG and then expand it to analyze severe acute respiratory syndrome corona virus 2 (SARS-CoV-2)-specific IgM and IgG, which is of great significance for the screening and diagnosis of COVID-19. In this study, the magnetic Fe(3)O(4) nanospheres coupled with mouse antihuman IgG (Ab1(IgG)) were used as an immune capture probe (Fe(3)O(4)@Ab1(IgG)) to capture and separate the target, and rabbit antihuman IgG (Ab2(IgG)) coupled with highly luminescent quantum dot nanobeads (QBs) as a fluorescence detection probe (QBs@Ab2(IgG)) was used to realize high sensitivity detection. After the formation of a sandwich immunocomplex, the fluorescence intensity of the precipitate after magnetic separation was measured at the excitation wavelength of 370 nm. Under optimal conditions, a wide linear range varying from 0.005 to 40 ng·mL(–1) was obtained for the detection of human IgG with a lower limit of detection at 4 pg·mL(–1) (S/N = 3). The recoveries of intra- and interassays were 90.0–101.9 and 96.0–106.6%, respectively, and the relative standard deviations were 6.3–10.2 and 2.6–10.5%, respectively. Furthermore, the proposed method was successfully demonstrated to detect human IgG in serum samples, and the detection results were not statistically different (P > 0.05) from commercial enzyme-linked immunosorbent assay kits. This method is sensitive, fast, and accurate, which could be expanded to detect the specific IgM and IgG antibodies against SARS-CoV-2.