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US 2025/0051758 A1Published application
FolA Selection Assay to Identify Strains with Increased Soluble Target Protein Expression
AssigneeAbSci LLC
InventorJia Liu
Life of this patent
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Abstract
The present disclosure provides compositions and methods for identifying cells or strains that produce soluble protein. The present disclosure further provides a growth/no growth selection strategy to identify E. coli strains in a mixed population (library) that produce higher soluble protein.
Claims27
A method of identifying a host cell capable of producing a soluble protein of interest, said method comprising the steps of:
- (a) preparing a population of host cells, wherein each host cell of the population comprises an expression construct capable of expressing a fusion protein comprising (i) a protein of interest and (ii) a selection protein;
- (b) incubating the host cells of (a) under conditions that allow expression of the fusion protein, wherein said conditions comprise a growth substance comprising at least 2 synergistic selection agents;
- and
- (c) visualizing host cells that are capable of growth;
- thereby identifying capable of producing a soluble protein of interest.
2 more elements
A method of identifying host cells that produce the highest amounts of a soluble protein of interest among a population of host cells, said method comprising the steps of:
- (a) preparing a population of host cells, wherein each host cell of the population comprises an expression construct capable of expressing a fusion protein comprising (i) a protein of interest and (ii) a selection protein;
- (b) incubating the host cells of (a) under conditions that allow expression of the fusion protein, wherein said conditions comprise a growth substance comprising at least 2 synergistic selection agents;
- and
- (c) visualizing host cells that are capable of growth;
- thereby identifying host cells that produce the highest amounts of a soluble protein of interest among a population of host cells.
2 more elements
Dependencies are inferred from the claim text.