Antibody preparation method, and antibody and antibody library thus prepared

摘要

Provided is a method for preparing antibodies against a protein of interest, through which highly specific antibodies against all proteins can be effectively and rapidly prepared with low cost, and the epitope to which the antibody is directed can be determined, so that a library covering epitopes on the surface of the proteins of interest and a library of antibodies against all the epitopes can be established. The antibodies are proved to be useful in detection, protein function investigation and antibody pharmaceuticals.

主权项

1. A method for preparing an antibody against a protein of interest, comprising:
(a) predicting and/or selecting peptide fragment located on the surface of the protein of interest, wherein said peptide fragment is a linear surface signature peptide and/or a conformational surface signature domain of the protein of interest; (b) synthesizing or expressing one or more of said peptide fragments; (c) using the product of step (b) to immunize an animal, optionally in combination with an adjuvant; (d) using lymphocyte from the immunized animal of step (c) to obtain antibodies; (e) using the peptide fragment of step (a) or said protein of interest in native conformation thereof to screen the antibodies obtained in step (d), so as to obtain an antibody library against said protein of interest, wherein said peptide fragment of step (a) is predicted or selected through the following process: (i) determining a surface peptide by calculating a parameter according to the sequence of the protein of interest, said parameter is selected from: solvent accessibility, disorder index, protein-protein interaction domain prediction, or any combination thereof; (ii) aligning the surface peptide determined in step (i) with the proteome of the species that the protein of interest is originated from, so as to select a specific peptide fragment of said protein of interest; (iii) aligning the surface peptide determined in step (i) with homologous proteins from other species, so as to screen a conservative sequence of said protein of interest.