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US 2022/0243222 A1Published application
Vectors and expression systems for producing recombinant proteins
AssigneeAmgen Inc.
InventorsPavan Ghattyvenkatakrishna, Hedieh Barkhordarian, Kristine Marie Daris, Charilyn Tejamo
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Abstract
Inventions disclosed herein relates to vectors and expression systems for producing heteromeric recombinant proteins such as monoclonal antibodies. Vectors and expression systems disclosed herein are based on the finding that the selectable marker glutamine synthetase can be divided into two fragments at selected amino acid positions of the glutamine synthetase polypeptide, and the two fragments can internet and/or associate to form a monomer and then a functional multimeric glutamine synthetase protein. Z
Claims48
A vector comprising:
- a) a first nucleic acid encoding a first polypeptide,
- b) a second nucleic acid encoding a first fragment of glutamine synthetase, wherein the transcription of the first nucleic acid is operably linked to the transcription of the second nucleic acid,
- c) a third nucleic acid encoding a third polypeptide, the third polypeptide is capable of associating with the first polypeptide to form a heteromeric complex, and
- d) a fourth nucleic acid encoding a second fragment of glutamine synthetase,
- wherein the transcription of the third nucleic acid is operably linked to the transcription of the fourth nucleic acid, and
- wherein the first fragment and the second fragment of glutamine synthetase associate to provide a selectable activity, and wherein the vector is capable of being transfected into mammalian cells and improving selection of transfected cells
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An expression system comprising:
- a) a first vector comprising a first nucleic acid encoding a first polypeptide, wherein the transcription of the first nucleic acid is operably linked to the transcription of a second nucleic acid encoding a first fragment of glutamine synthetase, and
- b) a second vector comprising a third nucleic acid encoding a third polypeptide, wherein the transcription of the third nucleic acid is operably linked to the transcription of a fourth nucleic acid encoding a second fragment of glutamine synthetase,
- wherein the first polypeptide is capable of associating with the third polypeptide to form a heteromeric complex,
- wherein the first and second fragments of glutamine synthetase associate to provide a selectable activity, and
- wherein the expression system is capable of being transfected into mammalian cells and improving selection of the cells.
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Dependencies are inferred from the claim text.