Highly avid magnetic bead capture: an efficient selection method for de novo protein engineering utilizing yeast surface display.

ABSTRACT: Protein engineering relies on the selective capture of members of a protein library with desired properties. Yeast surface display technology routinely enables as much as million-fold improvements in binding affinity by alternating rounds of diversification and flow cytometry-based selection. However, flow cytometry is not well suited for isolating de novo binding clones from naïve libraries due to limitations in the size of the population that can be analyzed, the minimum binding affinity of clones that can be reliably captured, the amount of target antigen required, and the likelihood of capturing artifactual binders to the reagents. Here, we demonstrate a method for capturing rare clones that maintains the advantages of yeast as the expression host, while avoiding the disadvantages of F

SUBMITTER: Ackerman M 

PROVIDER: S-EPMC2837102 | biostudies-literature | 2009 May-Jun

REPOSITORIES: biostudies-literature