Patent ID: 8945857
Filing Date: 2015-02-03
Classification: C07K,Y10S

Abstract:
1. A method of generating monoclonal antibodies specific for a desired antigen, comprising the steps: a) providing a sample from an animal that was exposed to the desired antigen 4-8 days prior, wherein the sample comprises a population of antibody-secreting cells that are secreting antibodies specific for the desired antigen; b) increasing a concentration of antibody-secreting cells in the sample by enriching for cells with general features characteristic of antibody-secreting cells; c) isolating a single antibody-secreting cell; d) cloning a first nucleic acid molecule that encodes the variable region of the heavy chain of the antibody made by the single antibody-secreting cell, and cloning a second nucleic acid molecule that encodes the variable region of the light chain of the antibody made by the single antibody-secreting cell; and e) transforming host cells with the cloned first and second nucleic acid molecules and expressing the first and second nucleic acid molecules that encode the variable regions or parts thereof of the heavy and light chains of the antibody made by the antibody-secreting cell; wherein steps d) and e) avoid the mixing of the first and second nucleic acid molecules with other nucleic acid molecules encoding heavy and/or light chain variable regions from different antibodies; and wherein the concentration of antibody-secreting cells in the sample by enriching for cells with general features characteristic of antibody-secreting cells according to b) is increased without using the desired antigen by a method selected from: