Humanized Collagen III Expression for Full-Length Mammalian Production

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Solution Overview

Problem

Current methods for producing recombinant collagen, particularly recombinant humanized collagen type III, face challenges such as truncated or fragmented proteins and differences in post-translational modifications when using Escherichia coli or yeast expression systems, which result in proteins that differ significantly from human collagen.

Innovation Solution

A recombinant humanized collagen type III alpha-1 (rhCol III α1) is fused with a green fluorescent protein (GFP) and expressed using an optimized expression vector pcDNA3.1 in human embryonic kidney cells, utilizing a secretion signal peptide and a tobacco etch virus protease digestion sequence to enhance full-length protein expression and promote cell migration.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If E. coli or yeast expression systems are used for recombinant collagen production, then large-scale expression is achieved, but the collagen becomes truncated, fragmented, or artificially modified

Engineering Contradiction:

Improvelarge-scale expressionVSAvoidcollagen structure accuracy

Core Design Contradiction:

ProductivityVSManufacturing precision

Solution Approach 1:

The patent changes the expression system from prokaryotic (E. coli) or simple eukaryotic (yeast) to mammalian cells (HEK293), which provide the appropriate cellular environment for proper collagen post-translational modifications and full-length protein synthesis, thereby resolving the contradiction between scale and precision

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent introduces an optimized expression vector pcDNA3.1 as an intermediary carrier that facilitates the transfer and proper expression of the collagen gene in mammalian cells, enabling both large-scale production and high structural fidelity

Inventive Principle:

Principle #24Intermediary (Mediator)

2Ease of manufacture

If animal-derived collagen is used, then mature extraction technology is available, but immunogenicity and limited sources occur

Engineering Contradiction:

Improveextraction technology maturityVSAvoidimmunogenicity

Core Design Contradiction:

Ease of manufactureVSObject-affected harmful factors

Solution Approach 1:

The patent creates a synthetic copy of human collagen type III alpha-1 chain through recombinant DNA technology, expressing the human collagen sequence in mammalian cells. This copy has identical primary structure to native human collagen but lacks the immunogenicity associated with animal-derived products, while being producible at scale

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent replaces the mechanical/chemical extraction process from animal tissues with a biological expression system using recombinant DNA technology in mammalian cells, substituting physical extraction with cellular synthesis to eliminate immunogenicity while maintaining manufacturability

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Data Source

PatentUS12570723B2Recombinant humanized collagen type III alpha-1, and expression vector and use thereof

Publication Date: 2026.03.10 SHANDONG D-NUTRIMEC BIOMEDICAL CO LTD

AI summary

A recombinant humanized collagen type III alpha-1 (rhCol III α1), and an expression vector and use thereof are provided, relating to the technical field of bioengineering. A sequence on positions 154 to 1,221 of a human type III collagen α chain is ligated to an optimized vector pcDNA3.1, a resulting optimized expression vector is transfected into a human embryonic kidney cell Expi293F, and then rhCol III α1 is secreted to form a fusion protein with a soluble green fluorescent protein (GFP). The expressed rhCol III α1 that forms the fusion protein with the GFP shows an activity of promoting the migration of BALB/c 3T3 cells, thus exhibiting a great application potential in the field of skin care.