DEEP Fusion Tags for Reliable Recombinant Protein Solubility
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing fusion tags used for recombinant protein expression in host cells, such as E. coli, are unreliable for producing a wide variety of proteins, leading to unpredictable expression yields and solubility issues, necessitating the testing of multiple tags for each protein of interest.
Innovation Solution
Development of De novo Expression Enhancer Protein (DEEP) fusion tags, which enhance expression and solubility of proteins over a broad pH range, facilitating their production and purification without the need for additional affinity tags.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If naturally occurring fusion tags are used for protein expression, then some proteins can be expressed, but expression reliability and solubility are unpredictable and vary by protein
Solution Approach 1:
The patent creates a universal fusion tag system where a single DEEP tag sequence can be applied to multiple different target proteins. The tag includes a multiple cloning site that allows insertion of various target protein sequences, making one tag design applicable to many different protein expression needs, thereby improving both reliability and versatility simultaneously
2Reliability
If multiple fusion tags are tested for each protein of interest, then expression optimization is achieved, but time and resources are consumed
Solution Approach 1:
The DEEP fusion tag is pre-designed with a multiple cloning site that accommodates various target proteins before actual expression experiments. This preliminary design allows researchers to directly use the optimized tag sequence for their specific protein without needing to test multiple different tags, thereby saving time while maintaining expression optimization
3Productivity
If existing fusion tags are used, then protein expression is facilitated, but solubility issues and degradation remain problems
Solution Approach 1:
The patent modifies the fusion tag sequence parameters by incorporating specific amino acid sequences (DEEP tag) that have been designed to improve protein solubility and stability. The tag includes specific residue patterns that prevent aggregation and degradation, thereby changing the physical-chemical parameters of the protein-tag fusion to achieve better solubility and stability while maintaining high production levels
Data Source
AI summary
The present invention relates, in certain embodiments, to a fusion protein comprising a DEEP fusion tag and a target protein (e.g., protein of interest). In various other embodiments, the invention provides methods of producing a fusion protein comprising a DEEP fusion tag and a target protein, constructs comprising a nucleotide sequence encoding a DEEP fusion tag and a cloning site for introducing a nucleotide sequence encoding a target protein, and kits comprising such DNA constructs.