Method of Screening Monoclonal Antibody Using SPR Technology

Abstract

The invention provides a method for screening monoclonal antibodies using SPR technology, which is to use an intermolecular interaction analyzer based on SPR technology to complete chip activation and coating; The pools are injected with different antibodies to bind the antibodies to the coated chip; when the analyte flows through to detect the antibody-antigen interaction, a single concentration of antigen is used as the analyte, and the buffer is not used as the background analyte; the raw data is directly analyzed Calculate the dissociation rate constant. Compared with the conventional method, the throughput of the method of the present invention is increased by 4 times, so that the SPR flux is suitable for the rapid screening requirements of the second screen; at the same time, the antigen consumption is reduced by one time, and the cost of the experiment is reduced; the dissociation rate constant produced by the method of the present invention The strong correlation with the results obtained by conventional methods can ensure the discovery of more antibody candidates with low MFI binding signal but slow dissociation rate.

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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