title: Utilization of Selenocysteine for Site-Specific Antibody Conjugation.
abstract: Site-specific conjugation methods are becoming increasingly important in building next-generation antibody-drug conjugates. We have developed a site-specific conjugation technology based on monoclonal antibodies with engineered selenocysteine (Sec) residues, named selenomabs. Here, we provide protocols for the engineering, expression, and purification of selenomabs in single-chain variable fragment (scFv)-Fc format. Methods for selective conjugation of selenomabs to selenol-reactive compounds and analytical characterization of selenomab conjugates are also included.
authors.affiliation: Department of Cancer Biology, The Scripps Research Institute, 130 Scripps Way #2C1, Jupiter, FL, 33458, USA.
authors.forename: Xiuling
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Cancer Biology, The Scripps Research Institute, 130 Scripps Way #2C1, Jupiter, FL, 33458, USA. crader@scripps.edu.|Department of Molecular Therapeutics, The Scripps Research Institute, 130 Scripps Way #2C1, Jupiter, FL, 33458, USA. crader@scripps.edu.
authors.forename: Christoph
authors.initials: C
authors.lastname: Rader
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D057809:HEK293 Cells; D006801:Humans; D018796:Immunoconjugates / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D015202:Protein Engineering; D017279:Selenocysteine / Q000737:chemistry / Q000378:metabolism*; D057127:Single-Chain Antibodies / Q000302:isolation & purification
chemical_list: D000911:Antibodies, Monoclonal; D018796:Immunoconjugates; D057127:Single-Chain Antibodies; D017279:Selenocysteine
keywords: Antibody-drug conjugates; Selenocysteine; Selenomab; Site-specific conjugation