Fructosyl-Peptide Oxidase

Product Overview

Diabetes poses a serious threat to human health. Glycated hemoglobin (HbA1c) reflects average blood glucose levels over the past 2-3 months and is essential for the diagnosis, treatment, and management of diabetes. Enzymatic HbA1c assays proceed in two steps: first, total hemoglobin concentration is measured; then, using a highly substrate-specific fructosyl peptide oxidase (FPOX) coupled with peroxidase, HbA1c concentration is determined via the Trinder reaction, from which the HbA1c percentage is calculated. Hzymes has developed a comprehensive solution spanning enzyme raw materials, chromogenic substrates, and assay reagents. Its proprietary patented fructosyl peptide oxidase (Patent No. CN202211041499.1) exhibits outstanding performance. Leveraging directed evolution and a high-throughput screening platform, the optimal FPOX mutant demonstrates a half-life at 45°C that is 4,000 times longer than that of the wild type, while its residual activity after acceleration remains tenfold higher than the wild type in HbA1c reagents. The dry powder boasts a specific activity of ≥4.0 U/mg, maintains stability across a pH range of 6.5-9.5, and is compatible with various reagent systems, with rigorously controlled levels of contaminating oxidase-related impurities. When used in conjunction with the protease (HH9958), peroxidase (HH2801), and DA67 chromogen (HH3701), the HbA1c reagent's calibrators and quality control materials are traceable to both IFCC and NGSP standards, delivering precise linearity across 3%-16% HbA1c. It also demonstrates excellent resistance to common interfering substances in clinical samples, enabling accurate measurement and compatibility with multiple fully automated biochemical analyzers. Hzymes independently develops and manufactures the complete suite of core raw materials for HbA1c assay reagents, ensuring full process control. Customized services for both raw materials and reagents are available, helping IVD manufacturers worldwide reduce costs and improve efficiency, fostering mutually beneficial outcomes.

Key Features

Ultra-Stable UDG: 4,000× Longer Half-Life at 45°C

Independently developed, with patent authorization CN202211041499.1.through targeted engineering, an outstanding mutant with significantly enhanced thermal stability was obtained, exhibiting a half-life at 45°C approximately 4,000-fold longer than that of the wild type.

10× Better Activity Retention After Accelerated Aging

After acceleration in the HbA1c assay reagent, the mutant exhibits tenfold the residual activity of the wild type, effectively ensuring the reagent's stability.

Exceptional Thermal Stability for Reliable Assay Performance

The thermal stability performance is significantly superior to that of similar products.

FPOX	Hzymes	VentorA	VentorB
Powder Specific activity (U/mg)	6.0	5.4	5.6
Optimal pH	8.0	8.0	7.5
Optimal temperature	45°C	45°C	45°C
Half-life at 47°C(min)	102	60	82

Applications

Core application: Diabetes testing-glycated hemoglobin assay. The accompanying protease (HH9958), peroxidase (HH2801), and DA-67 (HH3701) are used for the development and large-scale preparation of reagents for enzymatic detection of HbA1c.

Reaction Principle

The quinone imine formed in the reaction can be detected at a wavelength of 555 nm. One unit of enzyme activity is defined as the amount of enzyme required to catalyze the production of 1μmolof H2O2 per minute under conditions of pH 8.0 and 37°C.

Specification

CAS	109603-58-9
EC Number	EC 1.5.3
Appearance	Yellow amorphous lyophilized powder
Dry powder specific activity	≥4.0U/mg
Protein purity	≥90%(SDS-PAGE)
Protectants	Glycerin, trehalose
Contaminants	Catalase≤0.01%; ATPase≤0.005%; Glucose oxidase≤0.03%; Cholesterol oxidase≤0.003%
Optimal pH	pH6.5-7.5
pH stability	pH 6.5-9.5(25°C, 16 h)
Optimal temperature	37°C
Km	4.0×10^-3M (Fructosyl-Val-His)
Temperature stability	Below 40°C (pH8.0, 30 min)
Long-term stability	Stored at -20°C for 12 months, maintaining over 90% activity.
MW	60kDa(SDS-PAGE)
pI	6.4
Inhibitors	Hg²⁺, Pb²⁺

Available Formats

• HH1301-01 1kU

• HH1301-02 10kU

• HH1301-03 50kU

• HH1301-99 Customizable

Performance Data

Performance Summary of HbA1c (Enzymatic) Assay Reagent

Methodology	Enzymatic method
HbA1c reporting method	HbA1c(%) = HbA1c(μM) / Hb(μM)
Sample Type	EDTA Whole Blood Sample
Calibration Method	2-Point Linear
Consistency with HPLC	y = 1.017x -0.0143,R²= 0.9782
Linear range	3%~16% HbA1c (NGSP%)
Precision	Intra-batch≤3%
Precision	Inter-batch≤3%
Accuracy	≤3%
on-board stability	Stable for 30 days when stored at 2-8°C after opening.
Anti-interference	VitaminC≤50mg/dL, ≤±10%
	Chylomicrons≤200mg/dL, ≤±10%
	Uric acid≤5mg/dL ≤±10%
	Conjugated bilirubin≤15mg/dL, ≤±10%
Unconjugated bilirubin≤15mg/dL, ≤±10%

R² > 0.95 and <5% Average Deviation

Combining methodological rigor with the accuracy of internationally imported brands to meet clinical needs. The HbA1c enzymatic assay reagents from Hzymes demonstrate excellent correlation with results obtained by HPLC and with enzymatic assays from leading manufacturers, with correlation coefficients R²>0.95 and average deviations below 5%.

Stable Performance After 30 Days of Open-Vial Use

After 30 days of in-flight opening, performance remains as stable as ever.