Recombinant Human Collagen XVII Polypeptides for Low-Cost Production
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Solution Overview
Problem
Current methods for producing human-derived collagen face challenges such as high production costs, long cycles, and the risk of viral infection and sensitization from animal-derived collagen, which limits its application in biomedicine and tissue engineering due to coagulation risks and loss of biological activity.
Innovation Solution
Recombinant expression of human type 17 collagen polypeptides (C17A3, C17B3, and C17C1) using an optimized E. coli expression system, allowing for large-scale production with low costs and maintaining biological activity, stability, and cell adhesion properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Quantity of substance
If animal-derived collagen is used, then collagen products can be obtained, but the risk of viral infection and sensitization increases
Solution Approach 1:
The patent uses recombinant DNA technology to create a synthetic copy of human collagen type XVII in E. coli bacteria. The collagen gene is inserted into the bacterial genome, instructing them to produce identical human collagen polypeptides. This copying approach eliminates the need for animal tissues while maintaining human collagen's biological properties, thereby resolving the contradiction between production availability and safety risks.
2Reliability
If conventional recombinant expression methods are used for human-derived collagen, then human collagen can be produced, but production cost increases and production cycle lengthens
Solution Approach 1:
The patent optimizes multiple parameters of the E. coli expression system including codon optimization of the human collagen gene for bacterial expression, selection of appropriate promoters (T7 promoter), induction conditions (IPTG concentration and temperature), and cultivation parameters. These parameter optimizations enable high-level expression of human collagen type XVII in E. coli, achieving both biological activity and production efficiency simultaneously.
3Reliability
If conventional recombinant expression methods are used for human-derived collagen, then human collagen can be produced, but production cost increases
Solution Approach 1:
The patent utilizes E. coli bacteria as a disposable, inexpensive production platform. The bacteria can be rapidly cultured in simple media at low cost, and after expressing the collagen protein, the bacterial cells are discarded. This approach replaces expensive mammalian cell culture systems while maintaining the ability to produce biologically active human collagen, thereby reducing manufacturing costs.
4Reliability
If collagen with strong coagulation function is used, then collagen's natural function is maintained, but the risk of thrombosis increases
Solution Approach 1:
The patent extracts and expresses only the extracellular domain of human collagen type XVII (amino acids 63-1496), which contains the cell adhesion and structural functions. By excluding the transmembrane and intracellular domains that are involved in coagulation pathways, the recombinant polypeptide maintains collagen's beneficial structural and adhesive properties while eliminating the thrombosis risk associated with full-length collagen's coagulation function.
Data Source
AI summary
Provided is a polypeptide, a production method therefor and a use thereof. The polypeptide includes 63 to 1496 continuous amino acid residues of SEQ ID NO: 9, and includes a sequence represented by (A)m or is composed of the sequence represented by (A)m, wherein each A is an amino acid sequence selected from any one of those represented by SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, or a modified amino acid sequence or a sequence variant thereof; m is an integer between 1 and 10; and each A is the same or different and two adjacent As are directly connected by peptide bonds or connected by at least one amino acid residue, wherein the polypeptide has a cell adhesion activity, and a method of producing said polypeptide and use of said polypeptide.