Method of enabling pooled-library based nucleic acid constructs screening
AssigneeLeadxpro AG
InventorWassim Maxime Abdul Rahman
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Abstract
The invention relates to a method for generating in a single step a library of recombinant baculoviruses for screening for a protein of interest, starting from a pooled-library of poly-nucleotides each encoding a different protein. Insect cells are infected with a pooled-library of baculoviruses at a very low multiplicity of infection (MOI) leading to the situation where the majority of cells are infected by a single baculovirus. The pool of cells is then treated with a viral inhibitor which prevents secreted baculoviruses from penetrating other cells without affecting, either the capacity of the cell to express the target protein, nor intracellular viral replication and accumulation.
Claims16
A method for screening proteins and polypeptides to identify a protein or polypeptide of interest, which comprises the sequential steps of:
- i) generating a pooled-library of genes or genomic fragments cloned in baculovirus as a vector,
- (ii) transducing host cells with the pooled-library of baculovirus vectors at a multiplicity of infection (MOI) of less than 0.1,
- (iii) washing the host cells 2-4 hours post infection in a washing step,
- (iv) adding a viral inhibitor to the host cells,
- (v) culturing the cells for at least 2 days, so that the protein of interest is expressed in the host cells,
- wherein the viral inhibitor is selected from the group consisting of chloroquine, ammonium chloride, oseltamivir, or a neuraminidase inhibitor.
Dependencies are inferred from the claim text.