Modulating UPR Pathway Components for Antibody Secretion
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Solution Overview
Problem
Current methods for increasing protein secretion rates in cell lines are labor-intensive, time-consuming, and often result in low yields due to the rarity of high secreting cells and issues with protein misfolding in the endoplasmic reticulum, limiting the feasibility of protein purification.
Innovation Solution
Modulating the activity of unfolded protein response (UPR) pathway components in cells, such as BiP, XBP-1, and CHOP, through nucleic acid encoding siRNAs or small molecules, to enhance or decrease protein secretion rates by stabilizing or destabilizing the UPR pathway, thereby increasing or decreasing antibody secretion.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional methods of isolating high secreting cells through limiting serial dilution and screening are used, then high secreting cell lines can be identified, but the process is time-consuming and labor-intensive
Solution Approach 1:
The patent applies preliminary action by pre-modulating the UPR pathway components (such as overexpressing BiP, XBP-1, or CHOP) in cell lines before they are needed for protein production. This pre-conditioning of the cellular secretion machinery enables high secretion rates to be achieved directly, eliminating the need for time-consuming serial dilution and screening processes to identify high secreting cells.
2Reliability
If new cell lines are generated hoping they will be high secreting, then potentially high secreting lines can be obtained, but substantial effort is required to re-generate antibody producing cells and identify high secreting cells
Solution Approach 1:
The patent applies parameter changes by directly modifying the activity levels of UPR pathway components through various means such as overexpression, knockdown, or pharmacological modulation. By changing the functional state of these cellular components, the patent enables precise control over protein secretion rates, transforming the random search for high secreting cells into a targeted approach where secretion capacity is directly adjusted.
3Productivity
If protein misfolding occurs inside the endoplasmic reticulum, then secretion rate decreases, but the cellular secretion rate is an important parameter affecting production and purification
Solution Approach 1:
The patent applies the blessing in disguise principle by utilizing the UPR pathway, which is normally activated in response to ER stress and misfolding, and converting this harmful response into a beneficial mechanism for enhancing protein secretion. By deliberately modulating UPR components like BiP and XBP-1, the patent transforms the cell's stress response into a productive force that increases secretion capacity rather than merely compensating for misfolding.
Data Source
AI summary
Methods for altering the cellular secretion rate of a protein, such as an antibody and the altered cells produced by the method are disclosed. The methods and altered cells are useful for producing high levels of proteins for therapeutic, diagnostic or research purposes.