Why can recombinant factor C replace Limulus reagent for endotoxin detection?

1) The established utility of LAL;

In principle, the invention of the rFC method is based on the pathway understanding of the cascade reaction of endotoxin detection by the Limulus reagent method

The development of recombinant biotechnology will inevitably bring about the innovation of detection methods

Since Eli Lilly produced the first commercial recombinant human insulin, Humulin®, in 1982, more and more therapeutic macromolecules have been developed, bringing a new force to the biotechnology industry

As knowledge of the coagulation cascade continued to accumulate, scientists genetically engineered factor C, a serine protease that is extremely sensitive to endotoxin

The rFc method can cleave the fluorogenic substrate to generate fluorescence in one step, and feedback the endotoxin content according to the amount of substrate cleaved, which fully conforms to the 3R principle of experimental animals: reduce, replace, optimize, and the fourth "R" The principle, the reproducibility of endotoxin detection

In-depth understanding of the resistance faced by rFC detection methods in application

In the biomedical field, where pharmaceuticals and diagnostic products are highly regulated, it is important to use reliable alternative methods for LAL endotoxin detection, such as the rFc method, for testing of water, raw materials and process samples, as the use of alternative methods can save approximately 90% Limulus reagent was used

However, from RPT to LAL, from LAL to rFC, the replacement of endotoxin detection methods is based on the needs of industry development.

To date, the acceptance of the rFC detection method has been gradually increasing, and it is also a tried-and-tested supplement or alternative to LAL detection

The road ahead for rFC - regulatory and regulatory status (US, EU, China)

For many years, scientists have envisioned applying the results of the innate immunity study of horseshoe crabs to the biotechnology, pharmaceutical and human healthcare industries

From the perspective of resource availability, the continuous fishing of horseshoe crabs is also very unwise, and it will bring risks to the entire supply chain of pharmaceutical products
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Many scientists are also working to promote the use of synthetic rFCs instead of LALs to protect limulus from extinction, accelerating the adoption of rFCs as pharmacopoeial tests
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As the pharmaceutical industry evolves, more and more regulations permit rFC as an alternative to the LAL method (Table 1)
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More than 100 COVID-19 vaccine manufacturing companies are currently using rFC methods for quality control for efficient process production and quality assurance
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Lonza is the first manufacturer authorized to produce rFC factors, among which the PyroGene recombinant factor kit has been used by many pharmaceutical companies
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A global, multicenter study showed that endotoxin recovery from water and other test products using PyroGene was comparable to LAL-based methods
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The results of the kit validation were published in the Pharmacopoeia Forum Volume 36[1] (January-February 2010)
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For half a century since 1970, Lonza, as an early company developing endotoxin detection products, has provided global customers with a one-stop shop including bacterial endotoxin detection kits, endotoxin detection software, and endotoxin detection microplate readers.
The solution complies with FDA21CFR part 11 and EU EUGMP appendix 11 regulations, and has been used for the release and listing of a large number of foreign drugs to meet the large-scale endotoxin measurement needs of export-oriented/double-reported pharmaceutical companies
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