title: Coupling mammalian cell surface display with somatic hypermutation for the discovery and maturation of human antibodies.
abstract: A novel approach has been developed for the isolation and maturation of human antibodies that replicates key features of the adaptive immune system by coupling in vitro somatic hypermutation (SHM) with mammalian cell display. SHM is dependent on the action of the B cell specific enzyme, activation-induced cytidine deaminase (AID), and can be replicated in non-B cells through expression of recombinant AID. A library of human antibodies, based on germline V-gene segments with recombined human   regions was used to isolate low-affinity antibodies to human β nerve growth factor (hβNGF). These antibodies, initially naïve to SHM, were subjected to AID-directed SHM in vitro and selected using the same mammalian cell display system, as illustrated by the maturation of one of the antibodies to low pM K(D). This approach overcomes many of the previous limitations of mammalian cell display, enabling direct selection and maturation of antibodies as full-length, glycosylated IgGs.
authors.affiliation: AnaptysBio, Inc., 10421 Pacific Center Court, Suite 200, San Diego, CA 92121, USA. pbowers@anaptysbio.com
authors.forename: Peter M
authors.initials: PM
authors.lastname: Bowers
authors.forename: Robert A
authors.initials: RA
authors.lastname: Horlick
authors.forename: Tamlyn Y
authors.initials: TY
authors.lastname: Neben
authors.forename: Rachelle M
authors.initials: RM
authors.lastname: Toobian
authors.forename: Geoffrey L
authors.initials: GL
authors.lastname: Tomlinson
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Dalton
authors.forename: Heather A
authors.initials: HA
authors.lastname: Jones
authors.forename: Andy
authors.initials: A
authors.lastname: Chen
authors.forename: Laurence
authors.initials: L
authors.lastname: Altobell
authors.forename: Xue
authors.initials: X
authors.lastname: Zhang
authors.forename: John L
authors.initials: JL
authors.lastname: Macomber
authors.forename: Irina P
authors.initials: IP
authors.lastname: Krapf
authors.forename: Betty F
authors.initials: BF
authors.lastname: Wu
authors.forename: Audrey
authors.initials: A
authors.lastname: McConnell
authors.forename: Betty
authors.initials: B
authors.lastname: Chau
authors.forename: Trevin
authors.initials: T
authors.lastname: Holland
authors.forename: Ashley D
authors.initials: AD
authors.lastname: Berkebile
authors.forename: Steven S
authors.initials: SS
authors.lastname: Neben
authors.forename: William J
authors.initials: WJ
authors.lastname: Boyle
authors.forename: David J
authors.initials: DJ
authors.lastname: King
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry*; D001402:B-Lymphocytes / Q000276:immunology; D002462:Cell Membrane / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007075:Immunoglobulin M / Q000737:chemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation; D020932:Nerve Growth Factor / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D027041:Somatic Hypermutation, Immunoglobulin
chemical_list: D000906:Antibodies; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D020932:Nerve Growth Factor