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Bayer Aktiengesellschaft

Stichting Instituut Voor Dierhouderij En Diergezondheid

Stichting Institut Voor Dierhouderij En Diergezonheld

Dirk Franciscus Marinus Van De Wiel

Robertus Jacobus Maria Moormann

Dietmar Kretzdorn

Abraham Johannes De Smit

Erik Kees Hamann

Robert Hans Meloen

Petrus Antonius Van Rijn

The invention relates to a method of increasing protein expression in baculo vector virus expression systems. The invention provides a method to produce a recombinant protein in insect cell culture which comprises selecting a recombinant baculovirus expressing said protein, growing insect cells in growth medium in a culture vessel and infecting the cells with an inoculum of at least one baculovirus at a cell density of 1&#xd7;10to 5&#xd7;10cells/ml with an m.o.i of &#x3c;0.01. The invention also provides a method to produce recombinant pestivirus E2 or Eprotein or fragments thereof in insect cell culture characterized by a final concentration of the protein fragments in the growth medium at harvest of at least 100 &#x3bc;g/ml. The invention also provides a method of producing recombinant FSH, &#x3b1;-units and/or &#x3b2;-units, and complexes and fragments thereof, at a concentration in the growth medium at harvest of at least 15 &#x3bc;/ml.

E2 subunit vaccine comprising recombinant pestivirus E2 protein

The invention relates to a method to increase protein expression in baculo vector virus expression systems. The invention provides a method to produce a recombinant protein in insect-cell culture which comprises selecting a recombinant baculovirus expressing said protein, growing insect cells in growth medium in a culture vessel and infecting the cells with an inoculum of at least one baculovirus at a cell density of 1&times;10 to 5&times;10 cells/ml with an m.o.i of <0.01. The invention furthermore provides a method to produce recombinant pestivirus E2 or E protein or fragments thereof in insect cell culture characterized by a final concentration of said protein (fragments) in the growth medium at harvest of at least 100 &mgr;g/ml. The invention furthermore provides a method to produce recombinant follicle stimulating hormone, &agr;-units an/or &bgr;-units and complexes and fragments thereof, at a concentration in the growth medium at harvest of at least 15 &mgr;l.

Protein expression in baculovirus vector expression systems

The invention provides methods for the production of recombinant bovine Follicle Stimulating Hormone (bFSH) as well as vectors and cells for use in those methods. In particular the invention provides baculovirus based vectors which are capable of expression of bFSH in insect cells. bFSH is a heterodimeric protein belonging to a family of glycoprotein hormones which are produced in the pituitary or the placenta. It finds its use in many fertility related applications. Expression of bFSH in baculovirus/insect cell systems leads to a recombinant bFSH which has an unexpected high activity in a human FSH receptor assay and/or a bovine immature oocyte assay. The genes encoding the subunits of bFSH may be present on one baculovirus derived vector or on two or more vectors which are to be cotransfected.

Production of biologically active recombinant bovine follicle stimulation hormone (rec bFSH) in the baculovirus expression system

Growing community of inventors

Hardewijk, Netherlands

Dirk Franciscus Marinus Van De Wiel