Recombinant IL-15 Variant with C-Terminal Helix for High Expression
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Solution Overview
Problem
The development of native wild type IL-15 as a drug is hindered by low expression levels in prokaryotes and eukaryotes, difficulty in purification, and short half-life, leading to reduced protein yields and biological activity.
Innovation Solution
An IL-15 variant is created by adding a polypeptide of up to 30 amino acids to the carboxyl terminal of wild type human IL-15, incorporating basic hydrophilic amino acids and a linker to enhance stability and expression, resulting in increased yields and improved folding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If native wild type IL-15 is expressed in prokaryotes, then the protein can be produced, but the expression amount is low and the purification is difficult
Solution Approach 1:
The patent combines IL-15 with a carrier protein (such as albumin or immunoglobulin) to form a fusion protein. This merging approach increases the overall expression amount by leveraging the carrier protein's high expression levels and simplifies purification by using the carrier protein's inherent purification properties or tags.
Solution Approach 2:
The invention creates a composite protein structure by fusing IL-15 with a carrier protein. This composite approach allows the benefits of both proteins: IL-15 provides the desired biological activity while the carrier protein provides high expression levels and facilitates purification through its stable structure and available purification methods.
2Manufacturing precision
If multiple purification steps are performed to obtain high purity IL-15, then the purity increases, but the protein yield decreases and biological activity is reduced
Solution Approach 1:
By fusing IL-15 with a carrier protein that has high expression levels and stable structure, the invention reduces the number of purification steps required. The fusion protein can be purified in fewer steps while maintaining high purity, thereby preserving more of the protein's biological activity and increasing overall yield.
Solution Approach 2:
The invention changes the physical and chemical parameters of IL-15 by fusing it with a carrier protein. This modification alters the protein's solubility, stability, and interaction properties, enabling more efficient purification with fewer steps and better preservation of biological activity.
3Reliability
If native IL-15 is used for drug development, then the biological activity is present, but the half-life is short and the stability is reduced
Solution Approach 1:
The patent merges IL-15 with a carrier protein that has a long half-life and high stability (such as albumin or immunoglobulin). This fusion extends the half-life of IL-15 by leveraging the carrier protein's long circulation time and stability, while preserving IL-15's biological activity through the fusion design.
Solution Approach 2:
The invention creates a composite protein where IL-15 is fused with a stable carrier protein. This composite structure provides the benefits of both components: IL-15 retains its immunostimulatory biological activity while the carrier protein provides extended half-life and enhanced stability in vivo.
Data Source
AI summary
Provided is a recombinant interleukin-15 variant, wherein one or more amino acid residues are added, deleted and substituted at the end of an amino acid sequence of a wild type interleukin-15. Thus, the carboxyl terminal of interleukin-15 forms an amphipathic alpha helix structure, and the carboxyl terminal of the interleukin-15 variant has a more stable structure than the wild type interleukin-15, so that the interleukin-15 variant can be highly expressed in Escherichia coli, and is easier to be separated and purified. The expression quantity of the interleukin-15 variant is 10-20 times that of the wild type interleukin-15.