Sustainable rFC Endotoxin Assay Kit from China Suppliers: Reliable BET Solution for Pharmaceuticals and Medical Devices Factory
Technical Information on Recombinant Factor C (rFC) Fluorometric Assay for Bacterial Endotoxin Detection
Endotoxin LPS specifically binds to the LPS-recognition domain of rFC, triggering a conformational change that activates rFC’s serine protease activity. The intensity of AMC fluorescence is directly proportional to the endotoxin concentration in the sample, measurable via a fluorescence microplate reader.
Detects endotoxins at concentrations as low as 0.005 EU/mL, matching or exceeding traditional Limulus Amebocyte Lysate (LAL) assays.
Typically covers 0.005–50 EU/mL, enabling quantitative analysis across a broad concentration spectrum.
Exhibits high specificity for LPS; it is not activated by (1→3)-β-D-glucans, reducing false-positive results.
Requires controlled temperature (37±1°C), incubation time (60–120 minutes), and wavelengths of ~360 nm/~460 nm.
Prepare samples (e.g., pharmaceuticals, medical devices, biological products) via dilution or extraction to remove matrix interference.
Mix rFC reagent, fluorogenic substrate, and sample in a microplate well.
Incubate at 37°C to allow LPS-rFC binding, activation, and substrate cleavage.
Measure fluorescence intensity; calculate endotoxin concentration using a standard curve generated with known endotoxin standards (e.g., Escherichia coli O55:B5 LPS).
This assay combines sustainability (no horseshoe crab harvesting), reliability (low interference, high reproducibility), and efficiency, making it ideal for quality control in pharmaceutical and biotech industries.
Q: What is Recombinant Factor C (rFC)?
Recombinant Factor C (rFC) is an in vitro alternative for endotoxin detection. It is a recombinant protein engineered from the LPS-binding domain of horseshoe crab Factor C, eliminating the need to harvest natural horseshoe crab blood.
Q: How sensitive is the rFC fluorometric assay?
The assay is highly sensitive, with the ability to detect bacterial endotoxins at concentrations as low as 0.005 EU/mL. This matches or exceeds the performance of traditional Limulus Amebocyte Lysate (LAL) assays.
Q: Does the rFC assay suffer from interference from beta-glucans?
No. The rFC assay has high specificity for LPS and is not activated by (1→3)-β-D-glucans, which are common sources of interference and false-positive results in traditional LAL gel-clot assays.
Q: What are the standard incubation parameters for the assay?
The assay requires a controlled incubation temperature of 37±1°C for 60 to 120 minutes. Fluorescence intensity is subsequently measured at excitation/emission wavelengths of approximately 360 nm/460 nm.
Q: Is the rFC assay compliant with global pharmacopoeias?
Yes, the rFC fluorometric assay is recognized and compliant with major global pharmacopoeias, including USP <85> and EP 2.6.14, for quantitative bacterial endotoxin detection.