Recombinant Factor VII and IX Expression in Mammalian Cells
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for producing blood coagulation factors like Factor VIIa and Factor IX are inefficient, leading to difficulties in obtaining pure and large quantities, and are prone to viral contamination, making them unsuitable for consistent treatment of coagulation disorders.
Innovation Solution
The use of recombinant DNA technology to create DNA constructs encoding for Factor VII and Factor IX, which are then expressed in mammalian host cells, allowing for the production of proteins with similar biological activity, thereby eliminating viral contamination risks and ensuring a consistent source for treatment.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cryoprecipitate or plasma fractions are used for replacement therapy, then Factor VIIa and Factor IX can be obtained, but the production is inefficient and viral contamination risk remains
Solution Approach 1:
The patent uses recombinant DNA technology to create copies of the Factor VII and Factor IX genes in mammalian host cells. The genes are inserted into expression vectors that direct the host cells to produce identical copies of the coagulation factors, eliminating the need to extract them from human plasma and thereby eliminating viral contamination risks while enabling scalable production.
Solution Approach 2:
The patent introduces DNA constructs as an intermediary between the original human genes and the production system. These constructs include promoter regions, coding sequences, and regulatory elements that mediate the expression of Factor VIIa and Factor IX in mammalian host cells, enabling controlled and safe production without direct plasma extraction.
2Quantity of substance
If plasma pooling methods are used, then Factor VIIa and Factor IX can be produced, but large amounts of plasma are required and purity is difficult to achieve
Solution Approach 1:
The patent creates recombinant copies of the Factor VII and Factor IX genes in mammalian host cells using DNA technology. This allows production of large quantities of pure factors without needing to pool large amounts of plasma, as the host cells are engineered to specifically produce the desired coagulation factors with high purity.
Solution Approach 2:
The patent uses tissue-specific or inducible promoters in the DNA constructs to control where and when Factor VIIa and Factor IX are produced in the host cells. This localized expression control enables high purity production by directing synthesis to specific cell types or conditions, avoiding contamination from other plasma components.
3Reliability
If recombinant DNA technology is used to express Factor VII and Factor IX in mammalian cells, then pure and large quantities can be produced without viral contamination, but the complexity of the production system increases
Solution Approach 1:
The patent divides the Factor VII and Factor IX genes into functional segments including promoter regions, coding sequences, and regulatory elements. These segments are assembled into modular DNA constructs that can be independently optimized and combined, simplifying the overall design while maintaining high safety and consistency in factor production.
Data Source
AI summary
Methods are disclosed for producing proteins having biological activity for blood coagulation mediated by Factor VIIa or Factor IX. The proteins are produced by mammalian host cells which have been stably transfected with a DNA construct containing a nucleotide sequence which codes at least partially for either Factor VII or Factor IX. The nucleotide sequence comprises a first nucleotide sequence encoding a calcium binding domain, joined to a second nucleotide sequence positioned downstream of the first sequence. The second sequence encodes a catalytic domain for the serine protease activity of either Factor VIIa or Factor IX. The joined sequences code for proteins having substantially the same biological activity for blood coagulation as either Factor VIIa or Factor IX.