MULTI-PROTEASE METHOD Russian patent published in 2022 - IPC G01N33/68 C12Q1/37 

FIELD: biotechnology.

SUBSTANCE: present invention relates to a method for identifying an epitope on a protein, to which an antibody can bind. The method includes a stage of limited or partial protein proteolysis using one first protease or a combination of the first proteases and the next stage of proteolysis using one second protease or a combination of the second proteases, the specificity (specificities) of which differs from the specificity (specificities) of the first protease or proteases. Then, using mass spectrometry, peptides that have been released are analyzed to identify peptides, in which one end is cut by the first protease and the other end is cut by the second protease. One or more isolated epitopes are created, which are located in the region of the protein, containing or flanking the cutting site of the first protease. Antibodies directed at isolated epitopes are obtained. Using the specified antibodies, studies of one or more epitopes on the protein are conducted.

EFFECT: invention makes it possible to identify “missed” cutting sites, i.e., sites, which are cut by the first protease, but peptides are not released, and which are of potential interest as forming part of a suitable epitope.

17 cl, 15 dwg, 13 tbl, 4 ex