A homogeneous immunoassay for detection of the interaction between two tumor biomarkers of IGF1R-β and SOCS1.

Abstract

The current protein interaction method is time consuming and cumbersome or the instrument is expensive. A new method that is convenient, fast, and high throughput needs to be studied urgently. The purpose of this study was to establish a homogeneous immunoassay to detect the interaction between insulin-like growth factor-1 receptor-β (IGF1R-β) and suppressor of cytokine signaling 1 (SOCS1). The recombinant vectors IGF1R-β/pENTER and SOCS1/pENTER were constructed and transfected into 293T cells. Based on homogeneous immunoassay technology, we established a suitable method. The signal intensity in the 293T lysate that overexpressed IGF1R-β and SOCS1, respectively, was compared with the signal intensity in the simultaneous expression of IGF1R-β and SOCS1. The interaction between IGF1R-β and SOCS1 was verified in vitro. The detection system for the interaction between IGF1R-β and SOCS1 was established. Compared with other methods, homogeneous immunoassay has the advantages of being rapid and sensitive, having higher sensitivity, and easy to operate. The interaction between IGF1R-β and SOCS1 was tested to verify the feasibility of this method and prove its practicability and sensitivity. This new method can be used as a high-throughput platform for protein-protein interaction, with the advantages of trace detection, short detective time, and high detective sensitivity.