{ "ProductData": { "ID": "3.1.1.2.13.1", "ProductType": "Others", "BrandName": "α-2,3-Sialyltransferase, rec.", "ProductNameAddition": "human, expressed in HEK-293F, solution", "RegulatoryDisclaimer1": "For life science research only. Not for use in diagnostic procedures.", "DisclaimerGroup1": "LSR", "RegulatoryDisclaimer2": null, "DisclaimerGroup2": null, "RegulatoryDisclaimer3": null, "SampleType": [ ], "LicenseDisclaimers": [ { "Order": "78", "Identifier": "1403506419168", "LongText": "The sale of the Product does not exhaust or grant any rights in third party patents including patents of companies of the F. Hoffmann - La Roche AG group of companies, in particular, for the use of modified antibodies obtained by using the product." } ], "RelatedLinks": "", "Clone": "", "ControlTissue": [ "" ], "ISOtypes": "", "Species": [ "" ], "StainLocalization": [ "" ], "ProductNameGlobal": "alpha-2 3-Sialyltransferase rec." }, "ProductImageDetails": { "ImagePath": "https://pim-media.roche.com/Images/CB_IMAGE.png", "ImageType": "Image main" }, "Product2Taxonomy": { "Product2TaxonomyReferences": [ { "StructureSystemIdentifier": "Product_Grouping", "StructureSystemName": "Product Grouping", "StructureNodeStatus": "valid", "NodeName": "In vitro Glycoengineering", "NodeID": "02-0052" }, { "StructureSystemIdentifier": "OWP_Product_Types", "StructureSystemName": "Product Types", "StructureNodeStatus": "valid", "NodeName": "Glycosyltransferases", "NodeID": "70-025-00" }, { "StructureSystemIdentifier": "OWP_Organization", "StructureSystemName": "Organization", "StructureNodeStatus": "valid", "NodeName": "CustomBiotech and LifeScience", "NodeID": "01-01-01-07-00" }, { "StructureSystemIdentifier": "Product_Grouping", "StructureSystemName": "Product Grouping", "StructureNodeStatus": "valid", "NodeName": "Enzymes", "NodeID": "02-0014" }, { "StructureSystemIdentifier": "Product_Solutions", "StructureSystemName": "Product Solutions", "StructureNodeStatus": "valid", "NodeName": "CustomBiotech", "NodeID": "520" }, { "StructureSystemIdentifier": "Lab_Type", "StructureSystemName": "Lab Types", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "180-00" }, { "StructureSystemIdentifier": "Applications", "StructureSystemName": "Applications", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "99-00-00" }, { "StructureSystemIdentifier": "OWP_Techniques", "StructureSystemName": "Techniques", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "999-00" }, { "StructureSystemIdentifier": "Disease_Areas", "StructureSystemName": "Disease Areas", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "99-00-00" }, { "StructureSystemIdentifier": "Pathogens", "StructureSystemName": "Pathogens", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "99-00-00" }, { "StructureSystemIdentifier": "OWP_Family", "StructureSystemName": "Product Families", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "999" }, { "StructureSystemIdentifier": "Health_Topics", "StructureSystemName": "Health Topics", "StructureNodeStatus": "invalid", "NodeName": "", "NodeID": "99-00-00" } ] }, "Product2Materials": { "P2MaterialReferences": [ { "MaterialNum": "07429916103", "MaterialDescription": "alpha-2,3-Sialyltransferase, rec.", "RegisteredProductName": "alpha-2,3-Sialyltransferase, rec.", "GTIN": "07613336130766", "ProductCategoryText": "Reagents, kits", "OldMaterialNumber": "", "PackSizePIM360": "custom fill", "PackSizeDescPIM360": "Not Available", "MaterialAnnotation": "CustomBiotech product. Please contact your local representative.", "ReadyForUse": "", "OrderInformation": "Unit of measure is \"mg\"." } ] }, "Product2Products": { "Product2ProductReference": [ ] }, "ProductSpec": [ { "ProductSpecVariant": { "Chapters": [ { "Language": "en", "Value": "34.5 kDa", "Country": "XG", "Code": "Molecular Weight", "Name": "Molecular Weight" }, { "Language": "en", "Value": "Procedure and Assay Conditions

The optimal reaction conditions for sialylation of a target protein can be different for each application. Therefore, the presented protocol can only serve as a suggested starting protocol. Optimization of reaction conditions (e.g., with different enzyme-IgG-ratios) is recommended to determine the optimal conditions for each application.

1   Preparation of the reaction buffer, containing 6.1 mM CMP-NANA, 200 mM MES, pH 6.5.
•Dissolve 40 mg CMP-NANA and 390.4 mg MES in 6 ml of water.
•Adjust pH to 6.5.
•Adjust to a final volume of 10 ml with water.
2   Setting up the sialylation reaction, containing 500 μg CMP-NANA, 100 mg ST3Gal-6, 1 mg IgG.
Pipet the following solutions into a reaction tube:
•125 μl reaction buffer
•100 μl IgG (10 mg/ml, pH 6)
•18 μl α-2,3-Sialyltransferase
•7 μl water
3   Incubate the reaction at +37°C.
4   Stop the reaction by freezing the tube at -15 to -25°C.
5  
Analyze the samples

It is recommended to set up several identical reactions and stop them at different time points (e.g., 2 h, 8 h, 24 h) to plot the kinetics for each application.


Analytics

Samples were analyzed by electrospray ionization mass spectrometry and the content of G2+0SA, G2+1SA, and G2+2SA N-glycans was determined.
Instrument: Synapt G2 HDMS device (Waters, UK)
Software: MassLynx V 4.1
 ", "Country": "XG", "Code": "Protocols", "Name": "Protocols" }, { "Language": "en", "Value": "Unit of measure is \"mg\".", "Country": "XG", "Code": "CB - Order Information", "Name": "CB - Order Information" }, { "Language": "en", "Value": "As an example, highly galactosylated humanized monoclonal antibodies IgG1 and IgG4 are used for the kinetics.

Kinetics of α-2,3-Sialytransferase with IgG1
Maximal biantennary sialylation of IgG1 is observed after 24 hours (G2 + 2SA).


Kinetics of α-2,3-Sialytransferase with IgG4
Maximal biantennary sialylation of IgG4 is obtained after 8 hours (G2 + 2SA).", "Country": "XG", "Code": "Results", "Name": "Results" }, { "Language": "en", "Value": "For in vitro sialylation of all Galß1-4GlcNAc units on glycoproteins and complex molecules such as human monoclonal antibodies (MAB), use α-2,3-Sialyltransferase (ST3Gal-6) and CMP-N-Acetylneuraminic Acid (CMP-NANA).", "Country": "XG", "Code": "Applications (IFU)", "Name": "Applications (IFU)" }, { "Language": "en", "Value": "
Vial / Bottle Label Function / Description Content
1 α-2,3-Sialyltransferase rec. Enzyme
Storage buffer contains
50 mM MES, 200 mM NaCl, pH 6.4 at +4°C
Solution 
 
", "Country": "XG", "Code": "Content", "Name": "Content" }, { "Language": "en", "Value": "No animal-derived material is added in fermentation, purification and final formulation of this enzyme.", "Country": "XG", "Code": "Quality Control", "Name": "Quality Control" }, { "Language": "en", "Value": "Incubation of α-2,3-Sialyltansferase and CMP-NANA with a glycoprotein leads to increased sialylation of the glycoprotein.

As α-2,3-Sialyltansferase rec. adds sialic acid only to galactosylated glycostructures, a galactosylation of the glycoprotein in a previous step helps to achieve higher sialylation levels. Galactosylation can be achieved by incubation of the target protein with ß-1,4-Galactosyltransferase and UDP-Galactose.", "Country": "XG", "Code": "Applications", "Name": "Applications" }, { "Language": "en", "Value": "α-2,3-Sialyltransferase, rec. can be used for in vitro glycoengineering of glycoproteins, e.g. therapeutic proteins.", "Country": "XG", "Code": "Positioning", "Name": "Positioning" }, { "Language": "en", "Value": "Benefits of using in vitro glycoengineering to modify glycosylation of therapeutic proteins:
  • Time- and cost-saving generation of glycan variants to facilitate drug development.
  • Optimized glycosylation without compromising other CQAs or product yield.
  • Improved lot-to-lot consistency to reduce risk of product quality variation and resulting delays.
  • Generation of glycoprofiles which may not otherwise be generated.
  • Streamlined analytics in comparability studies.
", "Country": "XG", "Code": "Benefits", "Name": "Benefits" }, { "Language": "en", "Value": "α-2,3-Sialyltansferase, rec. is a highly active recombinant human glycosyltransferase, expressed in HEK-293F.", "Country": "XG", "Code": "Product Description", "Name": "Product Description" }, { "Language": "en", "Value": "Appearance: Clear, colorless to slightly colored solution.
Composition: 50 mM MES, 200 mM NaCl, pH 6.4 ± 0.1 at +4°C
Specific activity: ≥80 U/μg
Protein A280nm ( 1 mg/mL) = 0.877: 5.5 ± 0.5 mg/mL
Purity: ≥90%
Bioburden: ≤100 CFU/mL (in evaluation)
Endotoxin: ≤10 EU/mg (in evaluation)
Stability: At -15 to -25°C within specification range for 12 months.
", "Country": "XG", "Code": "Specification", "Name": "Specification" }, { "Language": "en", "Value": "Molar mass: 34.5 kDa by cDNA", "Country": "XG", "Code": "Properties", "Name": "Properties" } ] } } ] }

α-2,3-Sialyltransferase, rec.

human, expressed in HEK-293F, solution

logo

Ordering Information

Technical documents

for certificates, method sheets (instructions for use), safety data sheets and software downloads
error errorMessage
Sorry, we couldn't find the content you are looking for
Please try again later