title: Rapid optimization and prototyping for therapeutic antibody-like molecules.
abstract: Multispecific antibody-like molecules have the potential to advance the standard-of-care in many human diseases. The design of therapeutic molecules in this class, however, has proven to be difficult and, despite significant successes in preclinical research, only one trivalent antibody, catumaxomab, has demonstrated clinical utility. The challenge originates from the complexity of the design space where multiple parameters such as affinity, avidity, effector functions, and pharmaceutical properties need to be engineered in concurrent fashion to achieve the desired therapeutic efficacy. Here, we present a rapid prototyping approach that allows us to successfully optimize these parameters within one campaign cycle that includes modular design, yeast display of structure focused antibody libraries and high throughput biophysical profiling. We delineate this approach by presenting a design case study of MM-141, a tetravalent bispecific antibody targeting two compensatory signaling growth factor receptors: insulin-like growth factor 1 receptor (IGF-1R) and v-erb-b2 erythroblastic leukemia viral oncogene homolog 3 (ErbB3). A MM-141 proof-of-concept (POC) parent molecule did not meet initial design criteria due to modest bioactivity and poor stability properties. Using a combination of yeast display, structured-guided antibody design and library-scale thermal challenge assay, we discovered a diverse set of stable and active anti-IGF-1R and anti-ErbB3 single-chain variable fragments (scFvs). These optimized modules were reformatted to create a diverse set of full-length tetravalent bispecific antibodies. These re-engineered molecules achieved complete blockade of growth factor induced pro-survival signaling, were stable in serum, and had adequate activity and pharmaceutical properties for clinical development. We believe this approach can be readily applied to the optimization of other classes of bispecific or even multispecific antibody-like molecules.
authors: Merrimack Pharmaceuticals, Inc. Cambridge, MA, USA.
authors: Lihui
authors: L
authors: Xu
authors: Neeraj
authors: N
authors: Kohli
authors: Rachel
authors: R
authors: Rennard
authors: Yang
authors: Y
authors: Jiao
authors: Maja
authors: M
authors: Razlog
authors: Kathy
authors: K
authors: Zhang
authors: Jason
authors: J
authors: Baum
authors: Bryan
authors: B
authors: Johnson
authors: Jian
authors: J
authors: Tang
authors: Birgit
authors: B
authors: Schoeberl
authors: Jonathan
authors: J
authors: Fitzgerald
authors: Ulrik
authors: U
authors: Nielsen
authors: Alexey A
authors: AA
authors: Lugovskoy
mesh_terms: D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D016466:CHO Cells; D003412:Cricetulus; D015195:Drug Design; D015723:Gene Library; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D020893:Receptor, ErbB-3 / Q000276:immunology*; D017526:Receptor, IGF Type 1 / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use
chemical_list: D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D057127:Single-Chain Antibodies; C581292:ERBB3 protein, human; D020893:Receptor, ErbB-3; D017526:Receptor, IGF Type 1
keywords: ErbB3; IGF-1R; bispecific antibodies; high throughput screening; library; scFv; yeast display
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