LYTAC antibody conjugation

A 2 mg/ml solution of antibody was buffer exchanged into PBS using 7K Zeba size exclusion column. The antibody was reacted with 25 equiv. of NHS-(PEG)₄-Azide (20 mg/ml in DMSO), and the reaction was incubated overnight at rt. The reaction mixture was filtered using 7K Zeba size exclusion column to yield the conjugated antibody.

Tri-GalNAc-DBCO (100 equiv) was weighed into an Eppendorf tube and 2 mg/ml solution of Antibody-(PEG)₄-N₃ was added. The reaction was manually agitated until the mixture was homogeneous. The reaction mixture was allowed to incubate at rt in the dark for 3 days and filtered using 40K Zeba size exclusion column.