title: Engineering THIOMABs for site-specific conjugation of thiol-reactive linkers.
abstract: Antibody conjugates are used in many therapeutic and research applications and are generated by chemically linking a cysteine or lysine residue to potent chemotherapeutic drugs or other functional groups through a flexible linker. Recently, we have engineered THIOMABs (antibodies with engineered reactive cysteine residues) for site-specific conjugation and showed that these antibody conjugates display homogeneous labeling with optimal in vitro and in vivo characteristics. Here, we describe protocols for engineering, selection, and site-specific conjugation of THIOMABs with thiol-reactive linkers.
authors.affiliation: Genentech Inc., South San Francisco, CA, USA.
authors.forename: Sunil
authors.initials: S
authors.lastname: Bhakta
authors.forename: Helga
authors.initials: H
authors.lastname: Raab
authors.forename: Jagath R
authors.initials: JR
authors.lastname: Junutula
mesh_terms: D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry*; D057809:HEK293 Cells; D006801:Humans; D018796:Immunoconjugates / Q000737:chemistry* / Q000235:genetics*; D016297:Mutagenesis, Site-Directed; D015202:Protein Engineering / Q000379:methods*; D013379:Substrate Specificity
chemical_list: D018796:Immunoconjugates; D003545:Cysteine