Biological chips of surface plasma resonating biological sensor, preparation and application

Abstract

The invention relates to a cell surface receptor biochip based on a surface plasmon resonance biosensor, a preparation method and application thereof. The biochip is characterized in that the chip is provided with a gold surface coating at a glass substrate and the gold surface is fixed with a sephadex layer which is fixed with the monoclonal antibody of the Beta subunit of the receptors of anti para-insulin and surface receptors are fixed by antibody capture. The preparation method includes that the monoclonal antibody of the Beta subunit of the receptors of anti para-insulin adopts the method of antibody capture and is fixed on the surface of CM5 chip based on the surface plasmon resonance biosensor, so as to produce the protein chip of para-insulin receptors which is applicable to the mutual action between IGF-1R and IRS-1, SHC, PI3K or GRB2 and hopeful to be applied to screening cancer-fighting drugs.

Description

technical field [0001] The invention relates to a cell surface receptor biochip based on a surface plasmon resonance biosensor, a preparation method and an application thereof. The provided chip is mainly used for studying the interaction between insulin-like tyrosine kinase receptors and intracellular upstream signaling proteins Kinetics, and effects of kinase inhibitors on interactions. It belongs to the field of protein biochip. Background technique [0002] Traditional research methods for the interaction between cell surface receptors and their downstream signaling proteins include immunoprecipitation, Western blot and yeast two-hybrid. Immunoprecipitation and western blotting rely on the specificity of antibodies that bind to signaling proteins or receptors, and their sensitivity is low, which is not enough for the detection of weak interactions between proteins. Yeast two-hybrid has been used to study the interaction between insulin / insulin-like receptor (IR / IGF-1R)...

AI Technical Summary

Problems solved by technology

Immunoprecipitation and western blotting rely on the specificity of antibodies that bind to signaling proteins or receptors, and their sensitivity is low, which is not enough for the detection of weak interactions between proteins

Yeast two-hybrid has been used to study the interaction between insulin / insulin-like receptor (IR / IGF-1R) and its signaling protein. This method is more sensitive than immunoblotting, and the weak interaction between proteins can activate the transcription and translation of the reporter gene , but cannot give quantitative information, such as membrane receptors and extracellular ligands or intracellular receptor binding proteins and substrates due to the lack of corresponding methods to obtain the kinetic and thermodynamic characteristic constants of their interactions

Moreover, since the generation of the signal depends on the yeast transcription and translation system, the influence of the yeast system itself on the protein interaction cannot be ruled out

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