Recombinant Protein PTM Control via Host Modulators
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Solution Overview
Problem
The production of recombinant proteins in living cells is complicated by the difficulty in controlling post-translational modifications (PTMs), which is particularly challenging during biosimilar development and manufacturing process changes, as host proteins can introduce varying PTMs, making it hard to achieve a consistent PTM profile similar to reference products.
Innovation Solution
A method is developed to optimize cell culture conditions by adding modulators to the growth media to modulate the activity of host proteins responsible for PTMs, allowing for the production of recombinant proteins with target PTMs by measuring, setting target ranges, selecting appropriate modulators, and adjusting concentrations to match the desired PTM profile.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If host proteins are used to produce recombinant proteins, then cell productivity is improved, but PTM consistency deteriorates
Solution Approach 1:
Small molecule modulators are introduced as intermediaries to regulate host protein activity. These modulators bind to host proteins involved in PTM pathways (such as glycosylation enzymes) and fine-tune their activity levels, allowing the system to maintain both high productivity and consistent PTM profiles by controlling the intermediary molecules rather than eliminating the host proteins themselves
Solution Approach 2:
The invention changes the chemical parameters of the culture medium by adding specific small molecule modulators that alter the activity state of host proteins. By adjusting concentrations and types of modulators, the system can dynamically control PTM outcomes while maintaining productive protein expression, effectively decoupling productivity from PTM consistency
2Productivity
If process changes are made to improve productivity, then cell line performance is improved, but PTM profile consistency deteriorates
Solution Approach 1:
The invention applies preliminary action by pre-characterizing host protein activities and identifying appropriate small molecule modulators before process changes are implemented. This allows the development of a modulator addition strategy that can be applied to any process change scenario, ensuring PTM consistency is maintained even when productivity-enhancing process modifications are made
Solution Approach 2:
The system incorporates feedback mechanisms where PTM profiles are monitored and used to adjust modulator addition strategies. This feedback loop allows real-time or near-real-time correction of PTM variations that may arise from process changes, ensuring consistent output quality regardless of productivity optimization measures
3Manufacturing precision
If PTM control methods are implemented, then PTM consistency is improved, but process complexity increases
Solution Approach 1:
The invention employs small molecule modulators that are relatively simple, inexpensive compounds compared to complex biological reagents or engineered cell lines. These small molecules can be added directly to the culture medium and degraded or removed with the medium, avoiding the need for complex permanent genetic modifications or sophisticated equipment
Data Source
AI summary
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.