title: Yeast Display-Based Antibody Affinity Maturation Using Detergent-Solubilized Cell Lysates.
abstract: It is often desired to identify or engineer antibodies that target membrane proteins (MPs). However, due to their inherent insolubility in aqueous solutions, MPs are often incompatible with in vitro antibody discovery and optimization platforms. Recently, we adapted yeast display technology to accommodate detergent-solubilized cell lysates as sources of MP antigens. The following protocol details the incorporation of cell lysates into a kinetic screen designed to obtain antibodies with improved affinity via slowed dissociation from an MP antigen.
authors.affiliation: Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 3631 Engineering Hall, 1415 Engineering Drive, Madison, WI, 53706, USA.
authors.forename: Benjamin J
authors.initials: BJ
authors.lastname: Tillotson
authors.forename: Jason M
authors.initials: JM
authors.lastname: Lajoie
authors.forename: Eric V
authors.initials: EV
authors.lastname: Shusta
mesh_terms: D000915:Antibody Affinity; D000918:Antibody Specificity; D002457:Cell Extracts / Q000276:immunology*; D061505:Cell Surface Display Techniques / Q000379:methods*; D003902:Detergents / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000276:immunology*; D019151:Peptide Library; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D002457:Cell Extracts; D003902:Detergents; D008565:Membrane Proteins; D019151:Peptide Library; D057127:Single-Chain Antibodies