SmartEnzymes TransGLYCIT workflow

Transglycosylation of IgG

TransGLYCIT is a platform technology that enables efficient and site-specific human IgG glycan remodeling or conjugation. With TransGLYCIT, antibodies with defined and homogenous glycoforms or site-specific azide-activation are generated using fast and robust enzymatic workflows.

TransGLYCIT Remodeling generates a homogenous pool of antibodies carrying the G0, G1, G2 or G2S2 glycan profile with the core fucosylation intact, and TransGLYCIT Remodeling Afucosylated generates a homogenous pool of antibodies carrying the G0, G1, G2, G2S2 or Man5 glycan profile with core afucosylation. TransGLYCIT Azide Activation generates azide-activated antibodies prepared for conjugation with any label of choice using click-chemistry (DOL = 4).
 

  • Efficient and site-specific human IgG Fc N-glycan remodeling or conjugation
  • Prepares human IgG with homogenous glycoprofiles, or azide activates human IgG with a reliable degree of labeling (DOL) of 4
  • A rapid workflow – transglycosylation is completed within a few hours
Human IgG
Transglycosylation occurs
at the Fc N-glycan sites
3-6 hour workflow
G0, G1, G2, G2S2, Man5 or azide-activated glycoform is included

TransGLYCIT Remodeling

Glycan remodeling of human IgG with the G0, G1, G2 or G2S2 glycoform

TransGLYCIT Remodeling Afucosylated

Glycan remodeling of human IgG with the G0, G1, G2, G2S2 or Man5 glycoform, with core afucosylation

TransGLYCIT Azide Activation

Azide activation of human IgG1, IgG2 or IgG4

About TransGLYCIT

TransGLYCIT workflow

  1. Deglycosylation
    The Fc N-glycans are trimmed to the core GlcNAc using the IgG-specific GlycINATOR® Immobilized (EndoS2) enzyme that hydrolyses all Fc glycoforms, including high-mannose, hybrid, complex and bisecting glycans. For TransGLYCIT Remodeling, the degree of core fucosylation is the same as on the original IgG. It is possible to obtain afucosylated G0, G1, G2, G2S2 or Man5 glycoforms using TransGLYCIT Remodeling Afucosylated, where the FucosEXO™ 16 Immobilized enzyme hydrolyzes the α1,6-linked core fucose prior to the transglycosylation step.
  2. Transglycosylation
    The engineered glycosynthase TransINATOR™ catalyzes the transglycosylation reaction between the oxazoline-reactive G0, G1, G2, G2S2, Man5 or azide-activated glycoform and the core GlcNAc.

The transglycosylation of IgG is followed by an affinity purification step.
 
The enzymes in TransGLYCIT are expressed in E. coli and modified with His-tags.

TransGLYCIT Remodeling

Glycan remodeling of human IgG with the G0, G1, G2 or G2S2 glycoform.

TransGLYCIT Remodeling is a platform technology for efficient transglycosylation of native human IgG in a few hours. The robust workflow transglycosylates the antibody Fc N-glycans using enzymatic remodeling, resulting in a homogenous pool of antibodies carrying a defined glycoform. The technology is available in kits to generate antibodies carrying the G0, G1, G2 or G2S2 glycan profiles.

TransGLYCIT Remodeling contains sufficient material to transglycosylate 1 mg human IgG. The glycan-remodeled sample will have the same degree of fucosylation as the original molecule. If you wish to remove core fucosylation, we recommend using TransGLYCIT Remodeling Afucosylated.
 

  • Fast and efficient generation of human IgG with homogeneous glycoforms
  • Facilitates functional analysis of specific glycan structures
  • Reliable glycan remodeling achieved within a few hours

Available Products

TransGLYCIT Remodeling G0 1 mgGlycan remodeling of 1 mg human IgG with the G0 glycoformT1-G0F-0101,163.00Buy / Request a Quote
TransGLYCIT Remodeling G1 1 mgGlycan remodeling of 1 mg human IgG with the G1 glycoformT1-G1F-0101,163.00Buy / Request a Quote
TransGLYCIT Remodeling G2 1 mgGlycan remodeling of 1 mg human IgG with the G2 glycoformT1-G2F-0101,163.00Buy / Request a Quote
TransGLYCIT Remodeling G2S2 1 mgGlycan remodeling of 1 mg human IgG with the G2S2 glycoformT1-S2F-0101,163.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

TransGLYCIT Remodeling Citations

Unit Definition

TransGLYCIT transglycosylates ≥ 95% of 1 mg human IgG1 as measured by LC-MS.

Content and Storage

TransGLYCIT Remodeling contains:

  • GlycINATOR Immobilized Microspin
  • TransINATOR enzyme
  • Oxazoline glycoform G0, G1, G2 or G2S2
  • CaptureSelect™* Fc(ms) Microspin column for purification

TransGLYCIT Remodeling is shipped cold, and the content should be stored at different temperatures upon arrival.
 
Do not freeze the GlycINATOR Immobilized or the CaptureSelect™ columns!

Instructions

Safety Data Sheet

Certificate of Analysis

* Thermo Scientific™ CaptureSelect™ resin from Thermo Fisher Scientific Inc. and its subsidiaries. Thermo Fisher and CaptureSelect are trademarks of Thermo Fisher Scientific Inc. and its subsidiaries.

TransGLYCIT Remodeling Afucosylated

Glycan remodeling of human IgG with the G0, G1, G2, G2S2 or Man5 glycoform, with core afucosylation.

TransGLYCIT Remodeling Afucosylated is a platform technology for efficient transglycosylation of native human IgG in a few hours. The robust workflow transglycosylates the antibody Fc N-glycans using enzymatic remodeling, resulting in a homogenous pool of antibodies carrying a defined glycoform (G0, G1, G2, G2S2 or Man5) with core afucosylation.

TransGLYCIT Remodeling Afucosylated is provided in kits containing sufficient material to transglycosylate 1 mg human IgG.
 

  • Fast and efficient generation of human IgG with fully afucosylated homogeneous glycoforms
  • Complete removal of core fucose to investigate its impact in functional assays
  • Reliable glycan remodeling achieved within a few hours

Available Products

TransGLYCIT Remodeling Afucosylated G0 1 mgGlycan remodeling of 1 mg human IgG with the G0 glycoform and core afucosylationT1-G0A-0101,577.00Buy / Request a Quote
TransGLYCIT Remodeling Afucosylated G1 1 mgGlycan remodeling of 1 mg human IgG with the G1 glycoform and core afucosylationT1-G1A-0101,577.00Buy / Request a Quote
TransGLYCIT Remodeling Afucosylated G2 1 mgGlycan remodeling of 1 mg human IgG with the G2 glycoform and core afucosylationT1-G2A-0101,577.00Buy / Request a Quote
TransGLYCIT Remodeling Afucosylated G2S2 1 mgGlycan remodeling of 1 mg human IgG with the G2S2 glycoform and core afucosylationT1-S2A-0101,577.00Buy / Request a Quote
TransGLYCIT Remodeling Afucosylated Man5 1 mgGlycan remodeling of 1 mg human IgG with the Man5 glycoform and core afucosylationT1-M5A-0101,800.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

TransGLYCIT transglycosylates ≥ 95% of 1 mg human IgG1 as measured by LC-MS.

Content and Storage

TransGLYCIT Remodeling Afucosylated contains:

  • GlycINATOR/FucosEXO 16 Immobilized Microspin
  • TransINATOR enzyme
  • Oxazoline glycoform G0, G1, G2, G2S2 or Man5
  • CaptureSelect™* Fc(ms) Microspin column for purification

TransGLYCIT Remodeling Afucosylated is shipped cold, and the content should be stored at different temperatures upon arrival.
 
Do not freeze the GlycINATOR/FucosEXO 16 Immobilized or the CaptureSelect™ columns!

* Thermo Scientific™ CaptureSelect™ resin from Thermo Fisher Scientific Inc. and its subsidiaries. Thermo Fisher and CaptureSelect are trademarks of Thermo Fisher Scientific Inc. and its subsidiaries.

TransGLYCIT Azide Activation

Azide activation of human IgG1, IgG2 or IgG4.

TransGLYCIT Azide Activation enables efficient and site-specific conjugation at the Fc N-linked glycosylation sites of human IgG. Upon azide activation using TransGLYCIT Azide Activation, the antibody can be labeled with any alkyne-reactive click reagent of choice to a degree of labeling (DOL) of four labels per antibody (DOL = 4).

TransGLYCIT Azide Activation is available in kits containing sufficient material to azide activate 100 µg human IgG.

 

  • Facilitates site-specific custom conjugation of human IgG with a consistent degree of labeling (DOL) of 4
  • Higher DOL increases sensitivity for improved detection of low-level targets
  • Fc-specific labeling preserves antigen binding-capacity
  • Fast workflow for generation of azide-activated human IgG within a few hours

Available Products

TransGLYCIT Azide Activation hIgG1/4 100 𝜇gAzide activation of 100 𝜇g human IgG1 or IgG4T1-AZ1-001695.00Buy / Request a Quote
TransGLYCIT Azide Activation hIgG2 100 𝜇gAzide activation of 100 𝜇g human IgG2T1-AZ2-001695.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

TransGLYCIT Azide Activation transglycosylates ≥ 95% of 100 µg human IgG1 as measured by LC-MS.

Content and Storage

TransGLYCIT Azide Activation contains:

  • GlycINATOR Immobilized Microspin
  • TransINATOR enzyme
  • Oxazoline glycoform Azide
  • CaptureSelect™* Fc(ms) Microspin column for purification

TransGLYCIT Azide Activation is shipped cold, and the content should be stored at different temperatures upon arrival.
 
Do not freeze the GlycINATOR Immobilized or the CaptureSelect™ columns!

* Thermo Scientific™ CaptureSelect™ resin from Thermo Fisher Scientific Inc. and its subsidiaries. Thermo Fisher and CaptureSelect are trademarks of Thermo Fisher Scientific Inc. and its subsidiaries.

FAQ and Support

Popular FAQ

Unfortunately no, the enzyme requires trimming of the Fc-glycan using GlycINATOR to enable access to the core fucose substrate.

Yes, the transglycosylation reaction can be performed on all subclasses of human IgG. The reaction is somewhat slower on IgG2 and longer incubation times may be necessary to obtain over 95% transglycosylation.

GlycINATOR is an IgG specific endoglycosidase that hydrolyzes complex, hybrid and high mannonse type glycans on the conserved Fc site on IgG.

No, TransGLYCIT is based on IgG specific enzymes and will only transglycosylate IgG.

The TransGLYCIT platform is developed for transglycosylation of human IgG.

 

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