Host cell for producing a protein of interest
AssigneeLonza Ltd.
InventorsKatrien CLAES, James GRAHAM, Sylwia JOZWIAK, Christoph KIZIAK
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Abstract
A eukaryotic host cell engineered to produce a heterologous protein of interest (POI), which cell is genetically modified to reduce production of at least one of three different endogenous host cell protein (HCP), and its use in a method of producing the POI.
Claims15
A eukaryotic host cell engineered to produce a heterologous protein of interest (POI), which cell is genetically modified to reduce production of at least one endogenous host cell protein (HCP) selected from the group consisting of a first HCP (HCP1), a second HCP (HCP2), and a third HCP (HCP3), wherein
- a) HCP1 comprises the amino acid sequence identified as SEQ ID NO:1, if the host cell is Komagataella phaffii, or its homologous sequence that is endogenous to the host cell if of another species;
- b) HCP2 comprises the amino acid sequence identified as SEQ ID NO:3, if the host cell is Komagataella phaffii, or its homologous sequence that is endogenous to the host cell if of another species;
- and
- c) HCP3 comprises the amino acid sequence identified as SEQ ID NO:5, if the host cell is Komagataella phaffii, or its homologous sequence that is endogenous to the host cell if of another species.
A method for producing a protein of interest (POI) in a eukaryotic host cell, comprising the steps:
- i) genetically modifying the host cell to reduce production of at least one endogenous host cell protein (HCP) selected from the group consisting of a first HCP (HCP1), a second HCP (HCP2), and a third HCP (HCP3), wherein
- a) HCP1 comprises the amino acid sequence identified as SEQ ID NO:1, if the host cell is Komagataella phaffii, or its homologous sequence that is endogenous to the host cell if of another species;
- b) HCP2 comprises the amino acid sequence identified as SEQ ID NO:3, if the host cell is Komagataella phaffii, or its homologous sequence that is endogenous to the host cell if of another species;
- and
- c) HCP3 comprises the amino acid sequence identified as SEQ ID NO:5, if the host cell is Komagataella phaffii, or its homologous sequence that is endogenous to the host cell if of another species;
- ii) introducing into the host cell an expression cassette comprising one or more regulatory nucleic acid sequences operably linked to a nucleotide sequence encoding the POI;
- iii) culturing said host cell under conditions to produce said POI;
- and optionally
- iv) isolating said POI from the cell culture;
- and optionally
- v) purifying said POI.
Dependencies are inferred from the claim text.