Ultrasensitive detection of specific IgE based on nanomagnetic capture and separation with a AuNP-anti-IgE nanobioprobe for signal amplification

Abstract

The accurate detection of allergen specific IgE (sIgE) is fundamental in the diagnosis of allergic diseases. The present commercial platforms fail to meet the need for personalized diagnosis, due to the unsuitable allergen-fixation model and large amounts of serum consumption. In this work, we developed a nano-capturer Fe₃O₄@SiO₂-NTA with an enhanced signal by taking advantage of a AuNP-anti-IgE nanobioprobe for precise and highly sensitive quantification detection of sIgE in serum of allergic patients. The recombinant allergen was immobilized on Fe₃O₄@SiO₂-NTA through the interaction between its His-tag and Ni-NTA, which is more consistent with the real binding mode of allergens with sIgE in vivo than the present clinically used allergen-fixation methods. Numerous horseradish peroxidase (HRP)-labeled anti-IgE were modified onto one AuNP to detect the sIgE probed by Fe₃O₄@SiO₂-NTA@rCanf1. Once one anti-IgE binds to sIgE, other HRP-labeled anti-IgE modified on the same AuNP would all create signals, resulting in a significantly amplified chemiluminescence (CL) signal. Our results showed that this immunosensor could realize fast, accurate, low-cost and highly sensitive sIgE detection in serum samples. In vitro experiments demonstrated a 0.02 ng mL⁻¹ detection limit, which was lower than that of any standard analyzer used for allergy immunoassays. Furthermore, our method was utilized for the diagnosis of clinical samples. The results were in good agreement with those obtained by the clinical gold standard ImmunoCAP, with 1000 times less serum consumption than ImmunoCAP. Therefore, the presented immunosensor holds great promise to improve clinical sIgE quantitative detection and constitutes a potentially useful tool for clinical diagnosis and subsequent individual treatment of allergic diseases.