Genovis » GlyCLICK ADC MMAE

Description

GlyCLICK contains all reagents and materials needed to label IgG with GlyCLICK conjugation of DBCO-VS(GlcA)-PAB-MMAE.

Application	ADC Development, Antibody Conjugation
Product Group	GlyCLICK
Source/Origin	Cloned from S. pyogenes, Produced in E. coli (His-tagged), Engineered galactosyltransferase (Y289L)
Product Composition	Enzymatic components: Immobilized GlycINATOR (EndoS2) on agarose beads (20% EtOH), beta-1,4-galactosyltransferase (GalT)
Digestion Site	IgG Fc-glycans
Specificity	GlycINATOR hydrolyzes Beta-1,4-linked GlcNAc residues on all IgG Fc-glycoforms for site-specific azide activation using GalT and complete labeling using SPAAC
Substrates	Human IgG1, Human IgG2, Human IgG3, Human IgG4, Cow IgG, Fc-fusion proteins, Goat IgG, Horse IgG, Monkey IgG, Mouse IgG, Rabbit IgG, Rat IgG, Sheep IgG
Reaction Conditions	Deglycosylation (GlycINATOR): 120 min incubation at 37°C, Azide Activation (GalT): O/N incubation at 30°C, Labeling (SPAAC): O/N incubation at 25°C, *The protocol can vary depending on kit size/amount of IgG
Digestion Buffers	TBS (pH 7.4)
Buffer Compatibility	GlyCLICK steps are performed in TBS buffer, provided in the kit., For purification of azide activated Ab, TBS or PBS can be used., Sodium azide interferes with the click reaction and must be avoided., Avoid phophate buffers, not compatible with buffer additive (precipitation).
Additives/Co-factors	GalT requires buffer additive for activity., Buffer additive is included in the GlyCLICK kit.
Inhibitors	Denaturated IgG
Unit Definition	Components for antibody conjugation of up to 0.25-2 mg IgG
Label	MMAE with Cleavable Linker (Val-Ser(GlcA)-PAB)