Heparinase I for Diagnostics

Heparinase I rapidly and specifically neutralizes both unfractionated heparin as well as low molecular weight heparin, allowing baseline hemostasis to be monitored in the presence of these anticoagulants.

As a result, Heparinase I is incorporated into disposables from a number of leading diagnostic companies, and is in development for additional diagnostic uses

HEPARINASE I

HEPARINASE II

HEPARINASE III

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	DATA SHEET	FEB 2023, R.06
Heparinase I	Diagnostic Grade	Part No.
Synonyms	Heparinase; heparin lyase; heparin eliminase	50-008 50-009
Source	Flavobacterium heparinum (Recombinant)
EC Number	4.2.2.7
CAS Number	9025-39-2
Purity	≥ 95 % by reverse-phase HPLC analysis
Product Format	Heparinase I, PN 50-008 is presented in a phosphate buffered saline (PBS), pH 7.0; PN 50-009 in PBS, pH 7.0 containing a disaccharide as cryoprotectant. Supplied as frozen solution. No bovine serum albumin (BSA), glycerol or preservatives added.
Volume	Both PN 50-008 & PN 50-009 can be aliquoted from 10 µL to 900 mL per container as per customer’s request.
Storage & Shipping Information	Storage Temperature: -70 °C Transport Conditions: Product shipped on dry ice
Catalytic Reaction	The enzyme cleaves selectively (via an elimination mechanism) highly sulfated polysaccharide chains containing 1-4 linkages between hexosamines & O-sulfated iduronic acid residues. The reaction yields oligosaccharide products (mainly disaccharides) containing unsaturated uronic acids, which can be detected by UV spectroscopy at 232 nm. The enzyme also cleaves the antithrombin III binding pentasaccharide domain in the heparin molecule.
Substrate Specificity	Heparin; heparan sulfate (specific activity with heparin is approx. three times higher than with heparan sulfate).
Properties	• O-glycosylated at Ser-39 • Isoelectric point: 9.3 – 9.5 • Molecular weight: 42,508 Da • Calcium ion is a cofactor and an activator
Activity	• One International Unit (IU) is defined as the amount of enzyme that will liberate 1.0 µmole unsaturated oligosaccharides from porcine mucosal heparin per minute at 30°C & pH 7.0. (Activity depends on the assay temperature, the buffer, the source & the type of Heparin used). • One Unit (U) is also defined in other preparation as 1 U that liberates 0.1 µmol of unsaturated uronic acid per hour at 25°C and pH 7.5; 1 IU is equivalent to 600 U.

Intended Use, Reference & Precaution	• These products are inputs for further manufacture of medical diagnostic devices or for R&D use only & are not for therapeutic or other uses. • Refer to the respective lot-specific certificate of analysis for the actual activity & the shelf life. • Once thawed, aliquot as needed & freeze at -70oC to avoid multiple freeze-thaw cycles
Applications	• In vitro neutralization of heparin in blood & plasma samples before analysis. • Preparation of disaccharides of heparin & preparation of oligosaccharide libraries. • Measurement of heparin in blood & plasma using the in vitro thromboelastography (TEG) tests. • Coagulation & anticoagulation efficacy studies. • Production of low- & ultra-low molecular weight heparins (LMWH & ULMWH) from unfractionated heparin & immobilization of heparinase I for such use. • In-process, quality control, & compendial testing of heparins, heparan sulfate (HS), heparin- & HS-derived products. • Structural analyses, mass spectral analysis & characterization of heparin, heparan sulfate (HS), low molecular weight heparins, & synthetic heparin pentasaccharides & oligosaccharides. • Depolymerization of heparin, HS & chemically modified heparins, & molecular weight profiling of heparins. • Quantification of contaminants in heparin such as over-sulfated chondroitin sulfate & persulfonated heparin & quantification of process-related impurities. • Glycobiology & cancer biology research. • Identification of the biological properties of HS that depend on the integrity of the S-domains & determination of the spacing between S-domains. • In vitro host-pathogen interactions in viral infections, virusadhesion inhibition studies, virus-plaque inhibition assays, cell culture experiments, etc. • In vivo inhibition studies of neovascularization & proliferation of capillary endothelial cells. • Circumventing the inhibitory effects of heparin in PCR, RT-PCR, real-time RT-qPCR reaction & Western Blot. • In vitro histochemistry, immunohistochemistry, immunocytochemistry & flow cytometry, etc.