Fd Chain Gene 3' Terminus Modification for Antibody Secretion
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Solution Overview
Problem
Current methods for producing Fab-type antibodies using yeast as a host face challenges with low productivity and the risk of endoplasmic reticulum stress when using strong promoters to enhance production, leading to inefficient and costly processes.
Innovation Solution
Ligating a nucleotide sequence encoding 1 to 10 amino acid residues to the 3′-terminus of the Fd or L chain nucleotide sequence in yeast, allowing for improved secretion and production of Fab-type antibodies without the need for high-activity promoters, thereby reducing endoplasmic reticulum stress and production costs.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If a strong promoter with high activity is used to enhance antibody production, then the production amount of Fab-type antibody increases, but endoplasmic reticulum stress occurs and productivity decreases
Solution Approach 1:
The patent introduces a nucleotide sequence encoding a specific amino acid sequence (Asp-Lys-Thr-His-Thr) as an intermediary element at the 3'-terminus of the Fd or L chain gene. This intermediary sequence acts as a regulatory module that enhances antibody secretion without requiring high-activity promoters, thereby increasing production amount while maintaining productivity by avoiding endoplasmic reticulum stress.
2Quantity of substance
If a strong promoter with high activity is used to enhance antibody production, then the production amount of Fab-type antibody increases, but endoplasmic reticulum stress is given to cell mass
Solution Approach 1:
The patent introduces a nucleotide sequence encoding a specific amino acid sequence (Asp-Lys-Thr-His-Thr) as an intermediary element at the 3'-terminus of the Fd or L chain gene. This intermediary sequence acts as a regulatory module that enhances antibody secretion without requiring high-activity promoters, thereby increasing production amount while maintaining productivity by avoiding endoplasmic reticulum stress.
3Ease of manufacture
If conventional yeast host is used for protein production, then production cost decreases, but carbon source influences the product and productivity is low
Solution Approach 1:
The patent changes the genetic parameter of the yeast host by introducing a specific nucleotide sequence at the 3'-terminus of the antibody genes. This genetic modification enables conventional yeast to achieve high productivity without requiring special carbon sources, thereby maintaining low production costs while significantly improving antibody production levels.
Data Source
AI summary
It is an object of the present invention to provide a method for producing a low-molecular-weight antibody such as a Fab-type antibody, using yeast as a host, wherein the method is able to produce the low-molecular-weight antibody with high productivity. According to the present invention, there is provided a gene comprising a nucleotide sequence encoding an amino acid or an amino acid sequence capable of increasing the secretion amount of a Fab-type antibody at the 3′-terminus of a nucleotide sequence encoding the amino acid sequence of the Fd chain or L chain of an antibody.