title: IgG1 thioether bond formation in vivo.
abstract: During either production or storage, the LC214-HC220 disulfide in therapeutic antibodies can convert to a thioether bond. Here we report that a thioether forms at the same position on antibodies in vivo. An IgG1κ therapeutic antibody dosed in humans formed a thioether at this position at a rate of about 0.1%/day while circulating in blood. Thioether modifications were also found at this position in endogenous antibodies isolated from healthy human subjects, at levels consistent with this conversion rate. For both endogenous antibodies and recombinant antibodies studied in vivo, thioether conversion rates were faster for IgG1 antibodies containing λ light chains than those containing κ light chains. These light chain reaction rate differences were replicated in vitro. Additional mechanistic studies showed that base-catalyzed thioether formation through the light chain dehydrogenation was more preferred on antibodies with λ light chains, which may help explain the observed reaction rate differences.
authors.affiliation: Departments of Process and Product Development, Thousand Oaks, California 91320.
authors.forename: Qingchun
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Departments of Process and Product Development, Thousand Oaks, California 91320.
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Schenauer
authors.affiliation: Therapeutic Discovery, Amgen Inc., Thousand Oaks, California 91320.
authors.forename: John D
authors.initials: JD
authors.lastname: McCarter
authors.affiliation: Departments of Process and Product Development, Thousand Oaks, California 91320. Electronic address: gflynn@amgen.com.
authors.forename: Gregory C
authors.initials: GC
authors.lastname: Flynn
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D007145:Immunoglobulin kappa-Chains / Q000235:genetics / Q000378:metabolism*; D007146:Immunoglobulin lambda-Chains / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013440:Sulfides / Q000378:metabolism*
chemical_list: D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D011994:Recombinant Proteins; D013440:Sulfides
keywords: Antibodies; Disulfide; Drug Development; Light Chain; Mass Spectrometry (MS); Post Translational Modification