A paper-based ELISA for rapid sensitive determination of anaphylaxis-related MRGPRX2 in human peripheral blood

• Published in: Analytical biochemistry Vol. 633; p. 114392
• Main Authors: Ding, Yuanyuan, Li, Xiaoqian, Gao, Qingpeng, Dong, Xinyan, Kong, Liyun, Han, Shengli, Zhang, Tao, He, Langchong
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.11.2021
• Subjects: Anaphylaxis; Anaphylaxis - blood; Anaphylaxis - immunology; Covalent immobilization; Enzyme-Linked Immunosorbent Assay; Humans; MRGPRX2; Nerve Tissue Proteins - blood; Nerve Tissue Proteins - immunology; Paper-based ELISA; Receptors, G-Protein-Coupled - blood; Receptors, G-Protein-Coupled - immunology; Receptors, Neuropeptide - blood; Receptors, Neuropeptide - immunology; Paper-based ELISA; Anaphylaxis; Covalent immobilization; MRGPRX2
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2021.114392

Mas-related G-protein-coupled receptor X2 (MRGPRX2) has recently been reported to be associated with anaphylaxis. Detection of MRGPRX2 levels in human peripheral blood might serve as a powerful tool for predicting the predisposition of patients to anaphylactic reactions. For rapid measurement of MRGPRX2, we established a paper-based double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) using mouse monoclonal antibody and horseradish peroxidase (HRP)-labelled rabbit polyclonal antibody as capture antibody and detection antibody, respectively. We avoided chemical functionalization of the cellulose paper by introducing bovine serum albumin (BSA) to provide COOH and NH2 groups for covalent immobilization of the capture antibody. Through amide condensation, a two-layer immobilization strategy was applied with BSA-BSA and BSA-capture antibody networks as the first and second layers, respectively. This strategy improved the quantity, activity and stability of the immobilized antibody. We then established a paper-based ELISA to detect MRGPRX2 in human peripheral blood. Our method is less laborious, easier to implement, and more cost-effective than conventional ELISA, while offering similar sensitivity, specificity, and accuracy. Therefore, it could serve as an innovative clinical point-of-care diagnostic tool, especially in areas that lack advanced clinical equipment. [Display omitted] •A rapid and sensitive paper-based ELISA method was established to detect MRGPRX2.•A two-layer capture antibody immobilization strategy was applied.•Improved capture antibody immobilization effect and enhanced stability were achieved.•The established method is less laborious, easier to implement, and more cost-effective than conventional ELISA.