Heparinases II

Heparinase II, also known as Heparin Lyase II or Heparitinase II is catalytically active on both heparin and sulfated domains of heparan sulfate in an endolytic manner. It cleaves less selectively, via an elimination mechanism, sulfated polysaccharide chains containing 1-4 linkages between hexosamines and uronic acid residues (both iduronic and glucuronic acid residues) on the nonreducing end. The reaction yields oligosaccharide products (mainly disaccharides) containing unsaturated uronic acids which can be detected by UV spectroscopy at 232 nm. Complete depolymerization of polysaccharide chains is improved when Heparinase II is used in combination with Heparinase I and III for the disaccharide analysis.

The enzyme cleaves both heparin and heparan sulfate, with the heparan sulfate activity being 2-fold higher than the heparin activity.

Recombinant enzyme homologous to its native counterpart and expressed in the natural host, Pedobacter heparinus (formerly, Flavobacterium heparinum).

Heparinase II is O-glycosylated at Thr-134
Purity: ≥ 90 % by reversed phase HPLC analysis
IBEX liquid heparinase II has optimal activity and stability for up to 2.5 years when stored frozen at -70°C
IBEX lyophilized heparinase II has optimal activity and stability for up to two years when stored at 2-8^oC
Calcium is inhibitory

HEPARINASE I

HEPARINASE III

HEPARINASE I for DIAGNOSTICS

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Data Sheet - LIQUID


	DATA SHEET	FEB 2023, R. 07
Heparinase II	Research Grade	Part No.
Synonyms	Heparitinase II, Haparin lyase	50-011 (0.5 IU/vial) 50-011-001 (0.1 IU/vial) 50-011-002 (Bulk)
Source	Flavobacterium heparinum (recombinant)
EC Number	None assigned
CAS Number	149371-12-0
Purity	≥ 90 % by reverse-phase HPLC analysis
Product Format	Heparinase II is presented in a phosphate buffered saline pH 6.5 containing a disaccharide as cryoprotectant. Supplied as frozen solution. No bovine serum albumin (BSA), glycerol or preservatives added

Storage & Shipping Information	Storage Temperature: -70 °C Transport Condition: Product shipped on dry ice
Catalyzed Reaction	The enzyme cleaves, via an elimination mechanism, sulfated polysaccharide chains containing 1-4 linkages between hexosamines & uronic acid residues (both iduronic & glucuronic 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	The enzyme cleaves both heparin & heparan sulfate, with the heparan sulfate activity being about twice as high as the heparin activity.
Properties	• O-glycosylated at Thr-134 • Isoelectric point: 9.1 – 9.2 • Molecular weight: 85,765 Da • Calcium is inhibitory
Activity	One International Unit (IU) is defined as the amount of enzyme that will liberate 1.0 µmole unsaturated oligosaccharides from porcine mucosal heparin or heparan sulfate per minute at 30°C & pH 7.3. (Activity depends on the assay temperature, the buffer, the source & the type of heparan sulfate used).

Intended Use, Reference & Precaution	• These products are for in vitro R&D use only & 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 -70°C to avoid multiple freeze-thaw cycles.
Applications	• Production of low- & ultra-low molecular weight heparins (LMWH & ULMWH). • Characterization of heparin, heparan sulfate (HS) & LMWH, • Structural & mass spectral analyses & characterization of heparin, HS, LMWH, heparosan & Heparin-like GAGs. • Production & isolation of oligosaccharides with novel sequences of GlcNH3 + residues. • Depolymerization of heparin & HS, chemically modified heparins & molecular weight profiling of heparins, HS & chemically modified heparins. • In-process, quality control, & compendial testing of heparins, heparan sulfate (HS), heparin- & HS-derived products. • Quantification of contaminants & process-related impurities in heparin such as over-sulfated chondroitin sulfate & persulfonated heparin. • Glycobiology & cancer research. • In vitro host-pathogen interactions in viral infections, virus plaque inhibition assays, virus-adhesion inhibition studies, cell culture experiments, etc. • In vitro immunohistochemistry, immunocytochemistry & flow cytometry. • Circumventing the inhibitory effects of heparin in the PCR reaction.

Safety Data Sheets - LIQUID


	Safety Data Sheet - Heparinase II - Liquid
SECTION 1 – IDENTIFICATION
Release Date	May 2019 R. 01
Product Identity	Heparinase II, recombinant in sucrose, Research Grade
Part Number	50-011
Manufacturer	IBEX Pharmaceuticals Inc. 5485 Rue Paré, Suite 100 Montréal, Québec, H4P 1P7 Canada Tel : (514) 344-4004 Fax : (514) 344-8827
Product use	Research

SECTION 2 – HAZARD IDENTIFICATION
Emergency overview	Solution in water. Will not support burning. Not explosive.
Potential Health Effects
Skin	May cause skin irritation
Eyes	May cause irritation or burning sensation
Inhalation	May cause irritation to the respiratory tract
Swallowed	Not known to cause toxic effects
Injection	Not known to cause toxicity with acute exposure, but may produce toxicity with repeated injections.
Carcinogenicity	Not determined
SECTION 3 – COMPOSITION/INFORMATION ON INGREDIENTS
CAS No.	149371-12-0
EC No.	None assigned
Systematic name	Heparin lyase II
Synonyms	Heparinase II
Origin	Recombinant, non-pathogenic microorganism
Ingredients	No components need to be disclosed according to the applicable regulations.
SECTION 4 – FIRST AID MEASURES
Skin contact	Flush with copious amounts of water.
Eye contact	Rinse opened eye for several minutes under running water. Then consult a doctor.
Inhalation	Supply the victim with fresh air; consult doctor in case of complaints.
Swallowing	Rinse mouth with plenty of water.
On all of the above	Consult a doctor if symptoms develop.
SECTION 5 – FIRE FIGHTING MEASURES
Suitable extinguishing agents	Noncombustible.
Protective equipment	No special measures required. Prevent contact with skin and eyes
SECTION 6 – ACCIDENTAL RELEASE MEASURES
Person-related safety precautions	Prevent contact with skin and eyes. Ventilate area.
Measures for environmental protection	No special measures required.
Measures for cleaning/collecting	Spilled material should be carefully wiped up and disposed as biomedical waste as per applicable waste disposal regulations. Wash spill site after material pickup is complete.
SECTION 7 – HANDLING AND STORAGE
Handling
Information for safe handling	Prevent contact with skin and eyes and prevent inhalation and swallowing. No other special precautions are necessary if used correctly.
Information about protection against explosions and fires	No special measures required.
Storage
Requirements to be met by storerooms and receptacles	No special requirements except that product should be stored frozen (preferably at -70 °C).
Information about storage in one common storage facilit	Not required.
Further information about storage conditions	None
Storage class	None assigned
Class according to regulation on flammable liquids	Void
SECTION 8 – EXPOSURE CONTROLS / PERSONAL PROTECTION
General protective and hygienic measures	The usual precautionary measures for handling chemicals and potentially biohazardous materials should be followed. Use clothing sufficient to avoid skin contact. Safety shower and eye wash should be available in proximity.
Protection of hands	Use synthetic water resistant gloves.
Eye protection	Use safety glasses.
SECTION 9 – PHYSICAL AND CHEMICAL PROPERTIES
Physical form	Frozen, colorless, odorless aqueous solution. (Heparinase II is a protein)
Melting point	0 °C (melting point of aqueous solution)
Boiling point	Not applicable
Flash point	Not applicable
Flammability	Product is not flammable
Danger of explosion	Not determined
Solubility	Heparinase II and ingredients are water soluble
Organic solvents	0 %
Solids content	0 %
SECTION 10 – STABILITY AND REACTIVITY
Thermal decomposition / conditions to be avoided	No decomposition if used according to specifications.
Dangerous reactions	No dangerous reactions known.
Dangerous products of decomposition	No dangerous decomposition products known.
SECTION 11 – TOXICOLOGICAL INFORMATION
Effects of Acute Exposure	Low acute toxicity expected following inhalation, ingestion, injection or skin exposure. The potential eye irritation effects of Heparinase II are unknown and it should be considered to be potentially irritating to the eyes.
Effects of Chronic Exposure	Not determined for oral ingestion or skin exposure
Development Toxicity	Not determined
Reproductive Toxicity	Not determined
Carcinogenicity	Not determined
SECTION 12 – ECOLOGICAL INFORMATION
Toxicity	No data available
Persistence and degradability	No data available
Bioaccumulative potential	No data available
Mobility in soil	No data available
PBT and vPvB assessment	No data available
Other adverse effects	No data available
SECTION 13 – DISPOSAL INFORMATION
Waste Disposal	Incinerate or bury as a solid in a licensed facility. Do not discharge into waterways or sewer systems without proper authority.
Container Disposal	Dispose of in a licensed facility. Recommend crushing or other means to prevent unauthorized reuse.
SECTION 14 – TRANSPORT INFORMATION
DOT (US)	Not dangerous goods
IMDG	Not dangerous goods
IATA	Not dangerous goods
SECTION 15 – REGULATORY INFORMATION
Not a dangerous substance or mixture according to the Globally Harmonised System (GHS).
SECTION 16 – OTHER INFORMATION
This information is given without any warranty or representation. It is believed to be correct but does not claim to be all inclusive and shall be used only as a guide. IBEX Pharmaceuticals Inc. shall not be held liable for any damage resulting from handling or contact with the above product. It is offered solely for your consideration, investigation and verification.

Data Sheet - LYOPHILIZED


	DATA SHEET	FEB 2023, R02
Heparinase II Lyophilized	Research Grade	Part No.
Synonyms	Hepartinase II. Heparin Lyase II	60-018 (0.5 IU/vial) 60-019 (2 IU/vial)
Source	Flavobacterium heparinum (recombinant)
EC Number	None assigned
CAS Number	149371-12-0
Product Format	Heparinase II is presented in phosphate buffered saline pH 6.5 containing a disaccharide as lyoprotectant and lyophilized in a vacuum-sealed vial. No bovine serum albumin (BSA) or preservatives added.

Storage & Shipping Information	Storage Temperature: 2°C to 8°C Transport Condition: Shipped at ambient temperature

Catalytic Reaction	The enzyme cleaves (via an elimination mechanism) sulfated polysaccharide chains containing 1-4 linkages between hexosamines & uronic acid residues (both iduronic & glucuronic 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	The enzyme cleaves both heparin & heparan sulfate, with the heparan sulfate activity being about twice as high as the heparin activity.
Properties	• O-glycosylated at Thr-134 • Isoelectric point: 9.1 – 9.2 • Molecular weight: 85,765 Da • Calcium is inhibitor
Activity	One International Unit (IU) is defined as the amount of enzyme that will liberate 1.0 µmole unsaturated oligosaccharides from porcine mucosal heparin or heparan sulfate per minute at 30°C & pH 7.3. (Activity depends on the assay temperature, the buffer, the source & the type of heparan sulfate used).

Intended Use, Reference & Precautions	• These products are for in vitro R&D use only & not for therapeutic or other uses. • Refer to the lot-specific Certificate of Analysis (CoA) for the shelf life when the products are stored as lyophilized vials (without reconstitution) at 2 – 8°C and the actual activity post-reconstitution. • Reconstitute just before use. • DO NOT freeze the reconstituted enzyme.
Applications	• Production of low- & ultra-low molecular weight heparins (LMWH & ULMWH). • Characterization of heparin, heparan sulfate (HS) & LMWH, • Structural & mass spectral analyses & characterization of heparin, HS, LMWH, heparosan & Heparin-like GAGs. • Production & isolation of oligosaccharides with novel sequences of GlcNH3 + residues. • Depolymerization of heparin & HS, chemically modified heparins & molecular weight profiling of heparins, HS & chemically modified heparins. • In-process, quality control, & compendial testing of heparins, heparan sulfate (HS), heparin- & HS-derived products. • Quantification of contaminants & process-related impurities in heparin such as over-sulfated chondroitin sulfate & persulfonated heparin. • Glycobiology & cancer research. • In vitro host-pathogen interactions in viral infections, virus plaque inhibition assays, virus-adhesion inhibition studies, cell culture experiments, etc. • In vitro immunohistochemistry, immunocytochemistry & flow cytometry. • Circumventing the inhibitory effects of heparin in the PCR reaction

Safety Data Sheet - Lyophilized


	Safety Data Sheet - Heparinase II - Lyophilized
SECTION 1 – IDENTIFICATION
Release Date	February 28, 2018
Product Identity	Heparinase II, Lyophilized
Part Number	60-018, 60-019
Manufacturer	IBEX Pharmaceuticals Inc. 5485 Rue Paré, Suite 100 Montréal, Québec, H4P 1P7 Canada Tel : (514) 344-4004 Fax : (514) 344-8827
Product use	Research

SECTION 2 – HAZARD IDENTIFICATION
Emergency overview	Powder. Will not support burning. Not explosive
Potential Health Effects
Skin	May cause skin irritation
Eyes	May cause irritation or burning sensation
Inhalation	May cause irritation to the respiratory tract
Swallowed	Not known to cause toxic effects
Injection	Not known to cause toxicity with acute exposure, but may produce toxicity with repeated injections.
Carcinogenicity	Not determined
SECTION 3 – COMPOSITION/INFORMATION ON INGREDIENTS
CAS No.	149371-12-0
EC No.	None assigned
Systematic name	Heparin lyase II
Synonyms	Heparinase II
Origin	Recombinant, non-pathogenic microorganism
Ingredients	No components need to be disclosed according to the applicable regulations.
SECTION 4 – FIRST AID MEASURES
Skin contact	Flush with copious amounts of water.
Eye contact	Rinse opened eye for several minutes under running water. Then consult a doctor.
Inhalation	Supply the victim with fresh air; consult doctor in case of complaints.
Swallowing	Rinse mouth with plenty of water.
On all of the above	Consult a doctor if symptoms develop.
SECTION 5 – FIRE FIGHTING MEASURES
Suitable extinguishing agents	Noncombustible.
Protective equipment	No special measures required. Prevent contact with skin and eyes
SECTION 6 – ACCIDENTAL RELEASE MEASURES
Person-related safety precautions	Prevent contact with skin and eyes. Ventilate area.
Measures for environmental protection	No special measures required.
Measures for cleaning/collecting	Spilled material should be carefully wiped up and disposed as biomedical waste as per applicable waste disposal regulations. Wash spill site after material pickup is complete.
SECTION 7 – HANDLING AND STORAGE
Handling
Information for safe handling	Prevent contact with skin and eyes and prevent inhalation and swallowing. No other special precautions are necessary if used correctly.
Information about protection against explosions and fires	No special measures required.
Storage
Requirements to be met by storerooms and receptacles	No special requirements except that product should be stored refrigerated 2 - 8 °C
Information about storage in one common storage facilit	Not required.
Further information about storage conditions	None
Storage class	None assigned
Class according to regulation on flammable liquids	Void
SECTION 8 – EXPOSURE CONTROLS / PERSONAL PROTECTION
General protective and hygienic measures	The usual precautionary measures for handling chemicals and potentially biohazardous materials should be followed. Use clothing sufficient to avoid skin contact. Safety shower and eye wash should be available in proximity.
Protection of hands	Use synthetic water resistant gloves.
Eye protection	Use safety glasses.
SECTION 9 – PHYSICAL AND CHEMICAL PROPERTIES
Physical form	Lyophilized white cake in amber glass vial closed with cap covered with aluminum seal.
Melting point	Not applicable
Boiling point	Not applicable
Flash point	Not applicable
Flammability	Product is not flammable
Danger of explosion	Not determined
Solubility	Heparinase II and ingredients are water soluble
Organic solvents	0 %
Solids content	100 %
SECTION 10 – STABILITY AND REACTIVITY
Thermal decomposition / conditions to be avoided	No decomposition if used according to specifications.
Dangerous reactions	No dangerous reactions known.
Dangerous products of decomposition	No dangerous decomposition products known.
SECTION 11 – TOXICOLOGICAL INFORMATION
Effects of Acute Exposure	Low acute toxicity expected following inhalation, ingestion, injection or skin exposure. The potential eye irritation effects of Heparinase II are unknown and it should be considered to be potentially irritating to the eyes.
Effects of Chronic Exposure	Not determined for oral ingestion or skin exposure
Development Toxicity	Not determined
Reproductive Toxicity	Not determined
Carcinogenicity	Not determined
SECTION 12 – ECOLOGICAL INFORMATION
Toxicity	No data available
Persistence and degradability	No data available
Bioaccumulative potential	No data available
Mobility in soil	No data available
PBT and vPvB assessment	No data available
Other adverse effects	No data available
SECTION 13 – DISPOSAL INFORMATION
Waste Disposal	Incinerate or bury as a solid in a licensed facility. Do not discharge into waterways or sewer systems without proper authority.
Container Disposal	Dispose of in a licensed facility. Recommend crushing or other means to prevent unauthorized reuse.
SECTION 14 – TRANSPORT INFORMATION
DOT (US)	Not dangerous goods
IMDG	Not dangerous goods
IATA	Not dangerous goods
SECTION 15 – REGULATORY INFORMATION
Not a dangerous substance or mixture according to the Globally Harmonised System (GHS).
SECTION 16 – OTHER INFORMATION
This information is given without any warranty or representation. It is believed to be correct but does not claim to be all inclusive and shall be used only as a guide. IBEX Pharmaceuticals Inc. shall not be held liable for any damage resulting from handling or contact with the above product. It is offered solely for your consideration, investigation and verification.
Date of revision	February 28, 2018

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