Sustainable and Reliable BET Using Recombinant Factor C Fluorometric Assay
Catalog Number
Catalog No. |
Description |
Sensitivity EU/ml |
RFC100TA |
BETMAT™ rFC Endotoxin Assay Kit (Recombinant Factor C Fluorometric Method), 100 Tests/Kit |
0.01-10EU/ml |
RFC100TB |
0.005-5EU/ml |
Recombinant Factor C (rFC) Fluorescence-Based Bacterial Endotoxin Detection (BET) Product Features
Strong Sustainability:
By eliminating the use of horseshoe crab blood for lysate extraction, the recombinant Factor C is genetically engineered to protect endangered species and prevent overfishing. This sustainable approach aligns with global environmental and bioresource conservation initiatives, supporting green and compliant production practices.
High Detection Reliability:
With high specificity, rFC reacts exclusively with bacterial endotoxins and is not affected by impurities such as β-glucan, minimizing false positives and negatives. The assay achieves a sensitivity of up to 0.005 EU/mL and offers a broad linear detection range. Its repeatability and accuracy comply with international pharmacopoeia standards (USP, Eur.P), ensuring stable, consistent, and traceable results.
Wide Applicability and Convenience:
Compatible with various sample types, including vaccines, monoclonal antibodies, and medical device extracts, the rFC assay offers a simplified workflow suitable for automated testing systems and high-throughput analysis. The typical testing cycle is 1–2 hours, significantly improving laboratory efficiency.
High Safety and Stability:
Free from animal-derived components, the reagent eliminates contamination risks associated with traditional LAL reagents. It exhibits minimal batch-to-batch variation, flexible storage conditions, and a long shelf life, reducing operational costs and material waste.
Applications of Recombinant Factor C (rFC) Endotoxin Test Reagents
Recombinant Factor C (rFC) endotoxin detection technology is a sustainable, genetically engineered alternative to the traditional Limulus Amebocyte Lysate (LAL) test. It specifically recognizes the Gram-negative bacterial endotoxin lipopolysaccharide (LPS) with high sensitivity and interference resistance. It is widely used in pharmaceuticals, medical devices, biologics, laboratory testing, and environmental monitoring. In the pharmaceutical industry, rFC is used to control endotoxins in raw materials, injectables, and biopharmaceuticals to ensure drug safety. In the medical device sector, it can detect trace endotoxins on the surfaces of disposable and implantable products or in rinsates. In advanced therapies such as vaccines and cell and gene therapies, rFC can accurately measure endotoxins in complex samples, meeting global pharmacopoeia requirements. It is also used for testing cell culture systems, laboratory consumables, pharmaceutical water, and environmental samples. The rFC assay is not only precise and efficient, but also aligns with the concept of sustainable development, and is gradually becoming the new international standard for endotoxin testing.
Technical Information on Recombinant Factor C (rFC) Fluorometric Assay for Bacterial Endotoxin Detection
The rFC fluorometric assay is a horseshoe crab-free, in vitro method for quantitative bacterial endotoxin (lipopolysaccharide, LPS) detection, compliant with global pharmacopoeias (USP <85>, EP 2.6.14). Its core principle relies on recombinant Factor C (rFC)—a recombinant protein engineered from the LPS-binding domain of horseshoe crab Factor C—eliminating reliance on natural horseshoe crab blood and addressing sustainability concerns.
1. Assay Principle
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.
2. Key Technical Parameters
Sensitivity: Detects endotoxins at concentrations as low as 0.005 EU/mL, matching or exceeding traditional Limulus Amebocyte Lysate (LAL) assays.
Linear Range: Typically covers 0.005–50 EU/mL, enabling quantitative analysis across a broad concentration spectrum.
Specificity: Exhibits high specificity for LPS; it is not activated by (1→3)-β-D-glucans (a common interference in LAL gel-clot assays), reducing false-positive results.
Incubation Conditions: Requires controlled temperature (37±1°C) and incubation time (60–120 minutes), with fluorescence measured at excitation/emission wavelengths of ~360 nm/~460 nm.
3. Workflow Overview
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.