Antibody library screens using detergent-solubilized mammalian cell lysates as antigen sources.

ABSTRACT: High-throughput generation of antibodies against cellular components is currently a challenge in proteomics, therapeutic development and other biological applications. It is particularly challenging to raise antibodies that target membrane proteins due to their insolubility in aqueous solutions. To address these issues, a yeast display library of human single-chain antibody fragments (scFvs) was efficiently screened directly against detergent-solubilized and biotinylated lysates of a target cell line, thereby avoiding issues with membrane protein insolubility and eliminating the need for heterologous expression or purification of antigens. Antibody clones that specifically bind plasma membrane proteins or intracellular proteins were identified, depending on the biotinylation method applied

SUBMITTER: Cho YK 

PROVIDER: S-EPMC2920304 | biostudies-literature | 2010 Jul

REPOSITORIES: biostudies-literature