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Orthogonal Conjugation via Endoglycosidase and Microbial Transglutaminase Enables Efficient Synthesis of Dual-Payload Antibody-Drug Conjugates

ABSTRACT: Antibody drug conjugates (ADCs) are a rapidly growing field of therapeutics which combines the highly specific targeting ability of an antibody with a cytotoxic drug to deliver the payload in a targeted manner to cancer cells. Traditional ADC construction used stochastic conjugation methods resulting in heterogeneous products with subpopulations having mixed stability and pharmacokinetics. Consequently, recent efforts have favored site-specific conjugation methods that produce homogeneous ADCs with improved therapeutic indexes. Here we report a highly efficient site-specific conjugation method that combines an endoglycosidase and microbial transglutaminase to construct homogeneous and well defined antibody drug conjugates. We found an Endo-S2 treated IgG antibody, which conserves the inner-most GlcNAc residue at the conserved N-glycosylation site, is an excellent substrate for mTG conjugation activity. We further demonstrate an Fc glycan remodeling strategy with Endo-S2 can be used for loading of a second payload to construct a dual-drug ADC.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Peter Nemes

LAB HEAD: Peter Nemes

PROVIDER: PXD051902 | Pride | 2026-05-29

REPOSITORIES: Pride

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			Action	DRS
	20240310_AL_ADC_GNF_MMAE.msf	Msf
	20240310_AL_ADC_GNF_MMAE.raw	Raw
	20240328_AL01_GNF_Biotin.msf	Msf
	20240328_AL01_GNF_Biotin.raw	Raw
	20240328_AL02_MMAE_Exatecan.msf	Msf

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