Work

Yeast Display of Engineered Antibody Domains

Year: 2012

Type: book-chapter

Abstract: Yeast display is an efficient technology for selection of antibodies and other proteins with high affinity and thermal stability. Here, we describe a method for affinity maturation of engineered antibody domains (eAds) using yeast display. EAd yeast libraries of relatively large size (∼10⁹) were generated and subjected to alternating rounds of magnetic-activated cell sorting (MACS), fluorescent-activated cell sorting (FACS), and random mutagenesis. The highest affinity clones from the final round of maturation were identified and analyzed. We discuss extensively each key step, and provide detailed protocols and helpful notes.

Source: Methods in molecular biology

DOI: 10.1007/978-1-61779-921-1_5

Authors: Qi Zhao, Zhongyu Zhu, Dimiter S. Dimitrov

Institutions: National Institutes of Health, Frederick National Laboratory for Cancer Research, National Cancer Institute, Center for Cancer Research

Language: English

Topic: Monoclonal and Polyclonal Antibodies Research

Subfield: Radiology, Nuclear Medicine and Imaging

Field: Medicine

Domain: Health Sciences

Open Access status: green

Funders: Bill and Melinda Gates Foundation, National Institutes of Health, National Cancer Institute

Awards: Human Monoclonal Antibodies and Antibody-Guided Nanoparticles, Ligand-receptor-antibody interactions in transmembrane s