title: Removal of a C-terminal serine residue proximal to the inter-chain disulfide bond of a human IgG1 lambda light chain mediates enhanced antibody stability and antibody dependent cell-mediated cytotoxicity.
abstract: Optimization of biophysical properties is a critical success factor for the developability of monoclonal antibodies with potential therapeutic applications. The inter-domain disulfide bond between light chain (Lc) and heavy chain (Hc) in human IgG1 lends structural support for antibody scaffold stability, optimal antigen binding, and normal Fc function. Recently, human IgG1λ has been suggested to exhibit significantly greater susceptibility to reduction of the inter Lc-Hc disulfide bond relative to the same disulfide bond in human IgG1κ. To understand the molecular basis for this observed difference in stability, the sequence and structure of human IgG1λ and human IgG1κ were compared. Based on this Lc comparison, three single mutations were made in the λ Lc proximal to the cysteine residue, which forms a disulfide bond with the Hc. We determined that deletion of S214 (dS) improved resistance of the association between Lc and Hc to thermal stress. In addition, deletion of this terminal serine from the Lc of IgG1λ provided further benefit, including an increase in stability at elevated pH, increased yield from transient transfection, and improved in vitro antibody dependent cell-mediated cytotoxicity (ADCC). These observations support the conclusion that the presence of the terminal serine of the λ Lc creates a weaker inter-chain disulfide bond between the Lc and Hc, leading to slightly reduced stability and a potential compromise in IgG1λ function. Our data from a human IgG1λ provide a basis for further investigation of the effects of deleting terminal serine from λLc on the stability and function of other human IgG1λ antibodies.
authors.affiliation: Department of Antibody Technology; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Shen
authors.affiliation: Department of Antibody Technology; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Lin
authors.initials: L
authors.lastname: Zeng
authors.affiliation: Department of Antibody Technology; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Aiping
authors.initials: A
authors.lastname: Zhu
authors.affiliation: Department of Bioanalytical Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; Branchburg, NJ USA.
authors.forename: Tim
authors.initials: T
authors.lastname: Blanc
authors.affiliation: Department of Immunology; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Dipa
authors.initials: D
authors.lastname: Patel
authors.affiliation: Department of Oncology Translational Medicine; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Anthony
authors.initials: A
authors.lastname: Pennello
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Amtul
authors.initials: A
authors.lastname: Bari
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Stanley
authors.initials: S
authors.lastname: Ng
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Kris
authors.initials: K
authors.lastname: Persaud
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Yun Kenneth
authors.initials: YK
authors.lastname: Kang
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Balderes
authors.affiliation: Department of Oncology Translational Medicine; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: David
authors.initials: D
authors.lastname: Surguladze
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Sagit
authors.initials: S
authors.lastname: Hindi
authors.affiliation: Department of Bioanalytical Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; Branchburg, NJ USA.
authors.forename: Qinwei
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Department of BioProcess Sciences; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Dale L
authors.initials: DL
authors.lastname: Ludwig
authors.affiliation: Department of Antibody Technology; ImClone Systems, a wholly-owned subsidiary of Eli Lilly and Company; New York, NY USA.
authors.forename: Marshall
authors.initials: M
authors.lastname: Snavely
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000378:metabolism*; D000920:Antibody-Dependent Cell Cytotoxicity / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003545:Cysteine / Q000235:genetics; D057809:HEK293 Cells; D006358:Hot Temperature / Q000009:adverse effects; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics / Q000378:metabolism*; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D055550:Protein Stability; D012694:Serine / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007147:Immunoglobulin Light Chains; D012694:Serine; D003545:Cysteine
keywords: IgG1; IgG1λ; Lc; antibody; disulfide bond; lambda light chain; light chain; serine; stability and serine deletion