Recombinant Factor C Assay
Recombinant technology is a growing appeal in the pharmaceutical industry to help alleviate, and eventually replace, the need for animal resources for endotoxin testing purposes. Combining with horseshoe crab’s endotoxin-sensitive protein, Creative Biolabs provides a recombinant factor C (rFC) assay to detect the presence or absence of endotoxin in various test samples.
Principle of rFC
The rFC assay is the evolution of the LAL test and includes the rFC enzyme cloned from the horseshoe crab. The rFC is an endotoxin-sensitive synthetic protein that is cloned from factor C DNA. In this assay, when endotoxin binds to rFC, the activated synthetic rFC molecules that cleave a fluorescein substrate, amino-methylcoumarin result in the generation of a fluorogenic compound. The difference in fluorescence is proportional to endotoxin concentrations in the sample and the fluorescence is measured using 380/440 nm excitation/emission in the absence and presence of endotoxins. A comparative principle of LAL and rFC assay is shown in Fig.1.
Fig.1 Recombinant factor C pyrogen assay. A: Traditional LAL assay. B: The novel single-step activation of rFC. (Li, 2015)
Validation Timeline
A possible validation timeline is outlined below. The test with rFC method is performed in the step of validation of alternative method.
Fig.2 Validation timeline.
Assay Advantages
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No use of animal resources
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Through a single enzymatic step compared to LAL assays
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Provide a quantitative range of 5.0 EU/mL- 0.005 EU/mL in a single step
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Eliminating the dependence on nonspecific glycan
Creative Biolabs has experience in bacterial endotoxins testing for clients in the medical device industry. With a proven effective and compliant process, we provide the highest quality testing and fast turnaround time.For more detailed information, please feel free to contact us or directly send us an inquiry.
References
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Schneier, M.; et al. Current technologies to endotoxin detection and removal for biopharmaceutical purification. Biotechnology and Bioengineering. 2020, 117(8): 2588-2609.
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Li, P.; et al. Biotechnology efforts to conserve horseshoe crabs through the development of recombinant factor C-based endotoxin test. Changing Global Perspectives on Horseshoe Crab Biology. 2015.
For Research Use Only | Not For Clinical Use
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