Modified Inhibin Precursor Proteins for Efficient Cleavage
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Solution Overview
Problem
The production of recombinant inhibin A and B is challenging due to inefficient processing to their mature bioactive forms and co-production of activins, which complicates their therapeutic use, especially in conditions like osteoporosis related to reduced inhibin levels.
Innovation Solution
Development of mammalian inhibin analog precursor proteins with modified proprotein convertase cleavage sites and additional mutations to enhance processing efficiency, reduce activin production, and introduce tags for purification, allowing for the generation of bioactive inhibin A and B with minimal activin activity.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If recombinant inhibin production is attempted using standard methods, then inhibin can be produced, but processing efficiency to mature bioactive form is low and activin contamination occurs
Solution Approach 1:
The patent applies preliminary action by pre-modifying the precursor protein sequence before expression to include optimized proprotein convertase cleavage sites. This preliminary design ensures that once the protein is expressed, the cleavage process proceeds efficiently and accurately, preventing activin contamination from the outset rather than requiring extensive post-expression purification
Solution Approach 2:
The patent applies local quality by making specific localized modifications to the precursor protein sequence at the cleavage site regions. Rather than modifying the entire protein, targeted changes are made to the proprotein convertase recognition sequences to enhance cleavage efficiency and specificity, thereby improving mature inhibin yield without affecting other functional regions of the protein
2Quantity of substance
If standard recombinant expression is used, then inhibin precursor can be produced, but mature bioactive inhibin yield is low due to inefficient processing
Solution Approach 1:
The patent applies parameter changes by modifying the amino acid sequence parameters of the proprotein convertase cleavage sites. Specific substitutions are made to create optimal recognition sequences that enhance enzymatic cleavage efficiency, directly increasing the conversion rate from precursor to mature inhibin and thereby improving overall yield
3Productivity
If activin production is not prevented, then co-production occurs with inhibin, but this complicates therapeutic use
Solution Approach 1:
The patent applies the taking out principle by designing the precursor structure to separate inhibin and activin production pathways. By incorporating specific sequence modifications that direct proprotein convertase cleavage to produce inhibin heterodimers while preventing β-subunit homodimerization, the harmful activin byproduct is effectively extracted or prevented from forming alongside the desired inhibin product
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The modified precursor proteins result in higher yields of bioactive inhibin with reduced activin contamination, effectively addressing the limitations of existing recombinant production methods and providing a therapeutic potential for conditions associated with low inhibin levels.
Implementation Method 1
at least one proprotein convertase cleavage site in the α-subunit and/or β-subunit is modified by an amino acid substitution mutation to render it more efficiently cleaved by the proprotein convertase
Data Source
AI summary
The present invention relates to inhibin analogs, their method of production and their use in the treatment and prophylaxis of disease or conditions associated with reduced levels of inhibin and activin-mediated signaling.