ISOLATING CELLS EXPRESSING SECRETED PROTEINS

摘要

A method of detecting and isolating cells that produce a secreted protein of interest (POI), for example, an antibody, comprising: a) constructing a cell line transiently or stably expressing a cell surface capture molecule, which binds the POI, by transfecting the cell line with a nucleic acid that encodes such cell surface capture molecule; b) transfecting said cell simultaneously or subsequently with a second nucleic acid that encodes a POI wherein such POI is secreted; c) detecting the surface-displayed POI by contacting the cells with a detection molecule, which binds the POI; and d) isolating cells based on the detection molecule.

主权项

1. A method of detecting or isolating a eukaryotic cell that produces an antibody, comprising:
(a) providing a eukaryotic cell comprising (i) a nucleic acid encoding an antibody heavy chain, (ii) a nucleic acid encoding an antibody light chain, and (iii) a nucleic acid encoding a cell surface capture molecule, which comprises a membrane anchor and is capable of binding the antibody; (b) culturing the cell under conditions in which the antibody and cell surface capture molecule are expressed, and an antibody-cell surface capture molecule complex is formed intracellularly and displayed on the cell surface; (c) contacting the cell with a detection molecule, which binds to the antibody; and (d) detecting or isolating the cell.