title: Generation of human bispecific common light chain antibodies by combining animal immunization and yeast display.
abstract: Bispecific antibodies (bsAbs) pave the way for novel therapeutic modes of action along with potential benefits in several clinical applications. However, their generation remains challenging due to the necessity of correct pairings of two different heavy and light chains and related manufacturability issues. We describe a generic approach for the generation of fully human IgG-like bsAbs. For this, heavy chain repertoires from immunized transgenic rats were combined with either a randomly chosen common light chain or a light chain of an existing therapeutic antibody and screened for binders against tumor-related targets CEACAM5 and CEACAM6 by yeast surface display. bsAbs with subnanomolar affinities were identified, wherein each separate binding arm mediated specific binding to the respective antigen. Altogether, the described strategy represents a combination of in vivo immunization with an in vitro selection method, which allows for the integration of existing therapeutic antibodies into a bispecific format.
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Strasse 4, D-64287 Darmstadt, Germany.|Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Simon
authors.initials: S
authors.lastname: Krah
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Strasse 4, D-64287 Darmstadt, Germany.|Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Schröter
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Strasse 4, D-64287 Darmstadt, Germany.
authors.forename: Carla
authors.initials: C
authors.lastname: Eller
authors.affiliation: Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Laura
authors.initials: L
authors.lastname: Rhiel
authors.affiliation: Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Rasche
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Strasse 4, D-64287 Darmstadt, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Beck
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Strasse 4, D-64287 Darmstadt, Germany.|Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Sellmann
authors.affiliation: Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Günther
authors.affiliation: Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Toleikis
authors.affiliation: Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Hock
authors.affiliation: Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Alarich-Weiss-Strasse 4, D-64287 Darmstadt, Germany.
authors.forename: Harald
authors.initials: H
authors.lastname: Kolmar
authors.affiliation: Protein Engineering and Antibody Technologies, Merck KGaA, Frankfurter Strasse 250, D-64293 Darmstadt, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Becker
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D000912:Antibodies, Neoplasm / Q000096:biosynthesis / Q000235:genetics; D015703:Antigens, CD; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules / Q000037:antagonists & inhibitors; D058851:GPI-Linked Proteins / Q000037:antagonists & inhibitors; D006801:Humans; D051379:Mice; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D018033:Antibodies, Bispecific; D000912:Antibodies, Neoplasm; D015703:Antigens, CD; C527875:CEACAM5 protein, human; C504117:CEACAM6 protein, human; D002272:Carcinoembryonic Antigen; D015815:Cell Adhesion Molecules; D058851:GPI-Linked Proteins; D057127:Single-Chain Antibodies
keywords: antibody; bispecific antibody; common light chain; high-throughput screening; yeast surface display