Methods and compositions for displaying a polypeptide on a yeast cell surface
AssigneeOxyrane UK Limited
InventorsStefan Ryckaert, Guillaume Lerondel
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Abstract
Provided herein are methods and compositions for use in displaying a polypeptide (e.g., an antibody polypeptide or an antibody polypeptide fragment) on the surface of a yeast cell. Exemplary yeast that can be used in conjunction with various methods and compositions disclosed herein include those of the genus Yarrowia, e.g., Yarrowia lipolytica.
Claims83
A Yarrowia cell comprising an expression cassette comprising:
- a first promoter operably linked to a fusion sequence comprising a first nucleic acid sequence comprising a nucleotide sequence encoding a first antibody polypeptide, or a first antibody polypeptide fragment, fused in frame to an anchor polynucleotide sequence comprising a nucleotide sequence encoding an anchor polypeptide, wherein the first antibody polypeptide fragment comprises an antibody variable (V) region.
The cell of claims 1-15, further comprising a leader nucleic acid sequence comprising a nucleotide sequence encoding a leader polypeptide, wherein the leader nucleic acid sequence is fused in frame with and 5′ to the anchor polynucleotide sequence and the first nucleic acid sequence.
A method of expressing an antibody polypeptide or antibody polypeptide fragment in a Yarrowia cell, the method comprising:
- induction incubating a first Yarrowia cell,
- wherein the first Yarrowia cell comprises:
- (a) a first vector comprising a first promoter operably linked to a fusion sequence comprising a first antibody nucleic acid sequence comprising a nucleotide sequence encoding a first antibody polypeptide, or a first antibody polypeptide fragment, fused in frame to an anchor polynucleotide sequence comprising a nucleotide sequence encoding an anchor polypeptide, wherein the first antibody polypeptide fragment comprises an antibody variable (V) region;
- or
- (b) a first vector comprising a first promoter operably linked to a first antibody nucleic acid sequence comprising a nucleotide sequence encoding a first antibody polypeptide or a first antibody polypeptide fragment.
A method of selecting a Yarrowia cell comprising an antibody polypeptide, or antibody polypeptide fragment, that binds a target polypeptide, the method comprising:
- contacting a parent Yarrowia cell with the test polypeptide, wherein the parent Yarrowia cell displays on its surface a molecule comprising a first antibody polypeptide or a first antibody polypeptide fragment and wherein the parent Yarrowia cell comprises a first expression cassette comprising a first nucleic acid sequence comprising a nucleotide sequence encoding the first antibody polypeptide or the first antibody polypeptide fragment, wherein the first antibody polypeptide fragment comprises an antibody variable (V) region;
- and
- selecting the parent Yarrowia cell if the displayed molecule binds the target polypeptide.
Dependencies are inferred from the claim text.