Use of molecular chaperones for the enhanced production of secreted, recombinant proteins in mammalian cells
AssigneeBayer HealthCare LLC
InventorsSham-Yuen Chan, Hsinyi Yvette Tang, Yiwen Tao, Yongjian Wu, Ruth Kelly
Life of this patent
Estimating the term…
Abstract
The present invention relates to a method for increased production of a secreted, recombinant protein product through the introduction of molecular chaperones in a mammalian host cell. The present invention also relates to a mammalian host cell with enhanced expression of a secreted recombinant protein product by coexpressing at least one chaperone protein.
Claims12
A method for producing a secreted recombinant protein product comprising the steps of:
- culturing a mammalian host cell, said mammalian host cell having genetic material coding for expression of said recombinant protein product and transformed with at least one expression vector comprising DNA encoding Hsp70 chaperone protein, wherein the DNA encoding Hsp70 protein consists of the sequence SEQ ID NO:13;
- and
- recovering from the culture medium the recombinant protein product so produced and secreted.
A method for enhancing yield of recombinant Factor VIII or fragment thereof in a baby hamster kidney (BHK) cell line, wherein genetic material coding for expression of said recombinant Factor VIII or fragment thereof has been previously introduced into a first BHK cell line, said method comprising the steps of:
- inserting at least one Hsp70 chaperone protein expression vector into said first BHK cell line so as to form a modified BHK cell line, wherein the expression vector comprises DNA encoding Hsp70 chaperone protein, and wherein the DNA encoding Hsp70 chaperone protein consists of the sequence of SEQ ID NO:13;
- and
- selecting from said modified BHK cell line at least one second cell line exhibiting enhanced yield of the recombinant Factor VIII or fragment thereof.
A method for enhancing yield of a recombinant Factor VIII or fragment thereof in a BHK cell line comprising introducing genetic material coding for said Factor VIII or fragment thereof into a BHK cell line exhibiting enhanced Hsp70 chaperone protein expression, wherein the BHK cell line has been transformed with at least one expression vector comprising DNA encoding Hsp70, and wherein the Hsp70 chaperone protein consists of the sequence of SEQ ID NO:13.
Dependencies are inferred from the claim text.