Recombinant Vector BiP Fragment Minimization Foreign Peptide Sequences
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Solution Overview
Problem
Current methods for producing recombinant proteins face challenges in increasing production amounts and enhancing stability, particularly due to the addition of foreign peptide sequences during protein preparation, which can affect the efficacy and safety of biopharmaceuticals.
Innovation Solution
A recombinant vector is developed that includes a new BiP gene fragment, allowing for the sequential linkage of a polynucleotide encoding a target protein to minimize foreign peptide sequences and enhance expression levels, using promoters like 35S from cauliflower mosaic virus and transgenic organisms such as plants for protein production.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional methods are used to produce recombinant proteins, then production can be achieved, but the production amount is limited and foreign peptide sequences are added during preparation
Solution Approach 1:
The invention extracts and utilizes the BiP fragment (amino acids 1-240) from the endoplasmic reticulum, which naturally binds to recombinant proteins during their secretory pathway. By incorporating this fragment into the expression system, the foreign peptide sequences are minimized while maintaining protein production and purification capabilities.
Solution Approach 2:
The BiP fragment acts as an intermediary that facilitates the binding and purification of recombinant proteins. It serves as a mediator between the expression system and the target protein, enabling efficient purification through its natural protein-binding properties without requiring extensive foreign peptide tags.
2Ease of manufacture
If foreign peptide sequences are added during protein preparation, then purification can be facilitated, but the stability and efficacy of the biopharmaceutical is reduced
Solution Approach 1:
The invention extracts only the essential BiP fragment (amino acids 1-240) rather than using full-length BiP or large foreign peptide tags. This minimized foreign sequence approach maintains purification efficacy while reducing the impact on protein stability and efficacy.
Solution Approach 2:
The invention changes the parameter of foreign peptide sequence length by using a truncated BiP fragment (240 amino acids) instead of full-length BiP or conventional large tags. This parameter optimization maintains purification capability while minimizing negative effects on protein stability.
3Productivity
If conventional expression systems are used, then protein production can be achieved, but the use stability of the recombinant protein is reduced
Solution Approach 1:
The BiP fragment serves as a beneficial intermediary that is naturally present in the endoplasmic reticulum and binds to recombinant proteins during their secretory pathway. This natural interaction enhances protein stability without requiring large foreign peptide sequences that could interfere with protein function.
Data Source
AI summary
Provided is a recombinant vector including a new BiP gene fragment, and when a target protein is prepared using the recombinant vector of the subject matter, use stability can be enhanced and the production amount of target protein can also be increased by minimizing a foreign peptide sequence remaining in the target protein.