title: Remediating agitation-induced antibody aggregation by eradicating exposed hydrophobic motifs.
abstract: Therapeutic antibodies must encompass drug product suitable attributes to be commercially marketed. An undesirable antibody characteristic is the propensity to aggregate. Although there are computational algorithms that predict the propensity of a protein to aggregate from sequence information alone, few consider the relevance of the native structure. The Spatial Aggregation Propensity (SAP) algorithm developed by Chennamsetty et. al. incorporates structural and sequence information to identify motifs that contribute to protein aggregation. We have utilized the algorithm to design variants of a highly aggregation prone IgG2. All variants were tested in a variety of high-throughput, small-scale assays to assess the utility of the method described herein. Many variants exhibited improved aggregation stability whether induced by agitation or thermal stress while still retaining bioactivity.
authors.affiliation: a Department of Therapeutic Discovery; Amgen Inc. ; Thousand Oaks , CA USA.
authors.forename: Rutilio H
authors.initials: RH
authors.lastname: Clark
authors.forename: Ramil F
authors.initials: RF
authors.lastname: Latypov
authors.forename: Cyr
authors.initials: C
authors.lastname: De Imus
authors.forename: Jane
authors.initials: J
authors.lastname: Carter
authors.forename: Zien
authors.initials: Z
authors.lastname: Wilson
authors.forename: Kathy
authors.initials: K
authors.lastname: Manchulenko
authors.forename: Michael E
authors.initials: ME
authors.lastname: Brown
authors.forename: Randal R
authors.initials: RR
authors.lastname: Ketchem
mesh_terms: D000465:Algorithms; D020816:Amino Acid Motifs; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D000915:Antibody Affinity / Q000276:immunology; D002478:Cells, Cultured; D019295:Computational Biology / Q000379:methods; D057809:HEK293 Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000737:chemistry* / Q000276:immunology / Q000494:pharmacology; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D007694:Killer Cells, Natural / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008958:Models, Molecular; D066329:Protein Aggregates; D011485:Protein Binding / Q000276:immunology; D055503:Protein Multimerization; D055550:Protein Stability; D017434:Protein Structure, Tertiary; D013314:Stress, Mechanical
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D066329:Protein Aggregates; D007371:Interferon-gamma
keywords: CDR, complementary-determining regions; Fab, fragment antigen-binding; Fv, variable fragment; IgG, immunoglobulin G; RAP, relative aggregation propensity; SAP, spatial aggregation propensity; aggregation; agitation; antibody; computational; engineering; in silico; mAb, monoclonal antibody; modeling; stability; thermal