title: Identification of Novel Macropinocytosing Human Antibodies by Phage Display and High-Content Analysis.
abstract: Internalizing antibodies have great potential for the development of targeted therapeutics. Antibodies that internalize via the macropinocytosis pathway are particularly promising since macropinocytosis is capable of mediating rapid, bulk uptake and is selectively upregulated in many cancers. We hereby describe a method for identifying antibodies that internalize via macropinocytosis by screening phage-displayed single-chain antibody selection outputs with an automated fluorescent microscopy-based high-content analysis platform. Furthermore, this method can be similarly applied to other endocytic pathways if other fluorescent, pathway-specific, soluble markers are available.
authors.affiliation: UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, CA, United States.
authors.forename: K D
authors.initials: KD
authors.lastname: Ha
authors.affiliation: UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, CA, United States.
authors.forename: S M
authors.initials: SM
authors.lastname: Bidlingmaier
authors.affiliation: UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, CA, United States.
authors.forename: Y
authors.initials: Y
authors.lastname: Su
authors.affiliation: UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, CA, United States.
authors.forename: N-K
authors.initials: NK
authors.lastname: Lee
authors.affiliation: UCSF Helen Diller Family Comprehensive Cancer Center, University of California at San Francisco, San Francisco, CA, United States. Electronic address: bin.liu@ucsf.edu.
authors.forename: B
authors.initials: B
authors.lastname: Liu
mesh_terms: D000906:Antibodies / Q000276:immunology; D000915:Antibody Affinity / Q000502:physiology*; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology; D004705:Endocytosis / Q000502:physiology; D006801:Humans; D009369:Neoplasms / Q000276:immunology*; D019151:Peptide Library
chemical_list: D000906:Antibodies; D000951:Antigens, Neoplasm; D019151:Peptide Library
keywords: Cancer cell membrane dynamics; Cancer metabolism; Endocytosis pathway; High-content analysis; High-throughput screening; Human monoclonal antibodies; Macropinocytosis; Phage antibody display