Method for screening monoclonal antibodies by using SPR (Surface Plasmon Resonance) technology

Abstract

The invention provides a method for screening monoclonal antibodies by using the SPR technology. The method comprises steps of: completing a chip activation and a coating by using an intermolecular interaction analyzer based on the SPR technology; injecting different antibodies into each flow cell of each channel of the intermolecular interaction analyzer to bind the antibodies to the coated chip;using a single concentration of antigen as an analyte, without using the buffer as a background analyte, when the interaction of the antibody-antigen is detected through a flow analyte; and analyzingraw data directly to calculate a dissociation rate constant. The method of the invention increases the flux by 4 times compared with the conventional methods, so that the SPR flux is suitable for therapid screening requirement of the two sieves; at the same time, reduces the antigen consumption by one time; and reduces the experimental cost. The dissociation rate constants produced by the methodof the invention are highly correlated with the results obtained by conventional methods; so that the method can ensure the discovery of more antibody candidates with low MFI (Mean Fluorescence Intensity) binding signals but slow dissociation rates.

Description

technical field [0001] The invention relates to the field of biomacromolecule drugs, in particular to a method for screening monoclonal antibodies produced by hybridomas using SPR technology. Background technique [0002] Monoclonal antibodies are currently widely used to treat a variety of diseases, including autoimmune diseases and cancer. Since the US FDA approved the first antibody drug OKT3 in 1992, more than 80 therapeutic antibody drugs have been approved for marketing so far. The commonly used antibody discovery methods include hybridoma technology and phage display technology. When screening antibodies produced by hybridoma technology, ELISA is usually used to screen a large number (thousands) of antibodies first, and then select positive wells (200 to 300) with high OD values ​​and use FACS for screening. The second screen removes false positive molecules in ELISA. Most of the antibodies obtained at this time are antibodies that truly bind to the antigen. Final...

AI Technical Summary

Problems solved by technology

In this case, it takes too long to measure a large number of antibodies. For example, the second screening usually screens 250 antibodies, and the determination takes about 33 hours.

Since the antibody needs to be subcloned as soon as possible to prevent loss, the throughput of the Biacore 8K conventional experimental method is not enough to meet the requirements of the second screening, and the consumption of the antigen is also large

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