HIGH-THROUGHPUT ANALYSIS METHOD OF INTERACTION BETWEEN MATERIALS, USING PROTEIN FUSION DISPLAY TECHNIQUE

摘要

The present invention relates to a method for analyzing the interaction between a binding protein and a target material, including measuring the interaction between the binding protein and a target material using an interaction trapper (IT) cell. The IT cell has the binding protein displayed on the surface of intracellular inclusion bodies by expressing a fusion protein which forms active protein particles containing the binding protein, i.e., insoluble aggregates (inclusion bodies). The method includes increasing cell permeability which has no effect on the activity of the binding protein displayed on the cells and genetic information. Additionally, the present invention relates to a method for screening a library, further including a recovery by an individual cell unit. Additionally, the present invention relates to a cell and a cell library, which has a characteristic that can introduce a target material from the outside of a cell into the cells while conserving the activity of the binding protein displayed on the surface of intracellular inclusion bodies, and genetic information; and includes a construct containing a polynucleotide encoding the fusion protein, or the fusion protein such that a binding protein is displayed on inclusion bodies. The present invention has high screening efficiency due to the remarkably high signal-to-noise ratio compared to the known screening methods by 450 times via sensitively-labeling a target material interacting therewith through the overexpression of a binding protein. Also, the present invention provides high-throughput screening (HTS) which can readily isolate a single cell unit, thereby screening and isolating the interaction proteins in many libraries at ultra-high speed using a fluorescence microscope or flow cytometer. Additionally, the present invention can readily analyze the interaction of intracellular materials at high speed by introducing any foreign target material, which is difficult to be expressed in cells, into cells after the foreign target material is expressed outside of cells. The present invention can be used in various fields including developing antibodies/artificial antibodies, preparing a novel interacting protein, and analyzing and optimizing the interaction between a target protein and a drug candidate.