| 摘要 |
A method for optimizing post-translational modifications of recombinant proteins expressed in living cells is described. More particularly, a method for modulation of host proteins in living cells that control PTMs on recombinant proteins is described that has particularly useful applications in developing manufacturing process changes or in biosimilar development. The goal of this modulation is to produce a recipe for production of a recombinant protein in the new process or in the biosimilar that will produce a targeted PTM profile in the resulting protein product. In the method one or more modulators are selected, as from a modulator library, which affect the activity of host proteins. These modulators are added to media during production such that the resulting product matches the PTMs of the reference product. The ideal set of modulators and their concentrations are identified through a unique iterative process and the combined modulators and their concentrations constitute a recipe for growth media for the production of said recombinant protein. The methodology to obtain such a recipe described herein may then be used in many applications, such as optimizing new batches of recombinant protein drugs, developing biosimilar or bio-better drugs. |
| 主权项 |
1. A method for optimizing a growth medium for living cells that produce a recombinant protein having a target set of post-translational modifications, comprising:
A. measuring post-translational modifications on a reference, B. setting target post-translational ranges for the recombinant protein based on the measured post-translational modifications of the reference, C. providing a cell engineered to produce the recombinant protein, D. producing the recombinant protein from the cells engineered to express the recombinant protein in growth media containing a first set of modulators having a first set of concentrations, E. identifying the post-translational modifications on the recombinant protein and determining the differences between the identified post-translational modifications and the target post-translational modifications in B, F. developing a modified set of modulators wherein the modified set of modulators may include a different set of modulators, different concentrations of modulators, or a combination of different modulators and different concentrations of modulators, G. producing the recombinant protein from the cells engineered to express the recombinant protein in a growth media in the presence of the modified set of modulators, H. isolating the recombinant protein from growth media and comparing its post-translational modifications to the target set of post-translational modifications, I. repeating steps E, F, G and H until a set of modulators is identified that can be used to produce a recombinant protein having the target set of modulations. |
