Recombinant Human IL-2 Point Mutations for Secretion
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Solution Overview
Problem
Current methods for producing recombinant human IL-2 and its derived muteins are limited by low secretion levels and aggregation, which restricts their therapeutic potential.
Innovation Solution
Introduction of unique mutations, particularly at position 35 (K35E, K35D, and K35Q), in the genes encoding human IL-2 and its derived muteins, leading to increased secretion levels without affecting biological functions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If current methods are used for producing recombinant human IL-2 and its derived muteins, then production can be achieved, but secretion levels are low and aggregation occurs
Solution Approach 1:
The patent applies parameter changes by introducing specific point mutations at position 35 (K35E, K35D, K35Q) in the IL-2 gene sequence. These amino acid substitutions alter the protein's secretion parameters, increasing secretion levels by at least three times compared to wild-type IL-2 while simultaneously reducing aggregation tendencies, thus resolving the technical contradiction between productivity and harmful aggregation effects
2Productivity
If secretion levels are increased through mutations, then production efficiency improves, but biological functions must be maintained
Solution Approach 1:
The patent applies local quality by introducing mutations at a specific local position (position 35) in the IL-2 protein sequence. This localized modification at a single amino acid position optimizes secretion and reduces aggregation without disrupting the overall biological functionality of the cytokine, thereby achieving improved production efficiency while maintaining therapeutic activity
3Productivity
If point mutations are introduced to increase secretion, then secretion levels increase at least three times, but the gene sequence is modified
Solution Approach 1:
The patent applies parameter changes by making targeted modifications to the gene sequence at position 35, creating specific mutant variants (K35E, K35D, K35Q) that alter secretion parameters. The patent explicitly provides the modified gene sequences in Examples 1-3, demonstrating controlled sequence changes that achieve at least three-fold increase in secretion levels while maintaining the overall gene structure and biological function
Data Source
AI summary
The present invention relates to the field of Biotechnology, particularly to a method based on the introduction of a single mutation in the genes encoding the human IL-2 and the muteins derived thereof that results in increased secretion levels in different hosts without affecting their biological functions. In particular, these mutations are based on a non-conservative change in the amino acid located in position 35 in the primary sequence of human IL-2, preferably the substitutions are K35E, K35D and K35Q. Another object of the present invention are the expression systems used to obtain both the recombinant human IL-2 and the muteins derived thereof using the method described in this invention. The above-mentioned method is useful to improve the production efficiency of the recombinant human IL-2 and the muteins derived thereof both at laboratory and industrial scales. The proteins obtained using this method can be used for therapeutic purposes as well as in the in vitro expansion of T cells for adoptive transfer therapies.