title: Multiple Bacteriophage Selection Strategies for Improved Affinity of a Peptide Targeting ERBB2.
abstract: Due to the heterogeneity of ERBB2-expression between tumors and over the course of treatment, a non-invasive molecular imaging agent is needed to accurately detect overall ERBB2 status. Peptides are a highly advantageous platform for molecular imaging, since they have excellent tumor penetration and rapid pharmacokinetics. One limitation of peptides however, is their traditionally low target affinity, and consequently, tumor uptake. The peptide KCCYSL was previously selected from a bacteriophage (phage) display library to bind ERBB2 and did so with moderate affinity of 295 nM. In order to enhance tumor uptake and clinical utility of the peptide, a novel phage microlibrary was created by flanking the parent sequence with random amino acids, followed by reselection using parallel strategies for high affinity and specific ERBB2 binding in an attempt to affinity maturate the peptide. One limitation of traditional phage display selections is difficulty in releasing the highest affinity phages from the target by incubation of acidic buffer. In an attempt to recover high affinity second-generation peptides from the ERBB2 microlibrary, two elution strategies, sonication and target elution, were undertaken. Sonication resulted in an approximately 50-fold enhancement in recovered phage per round of selection in comparison to target elution. Despite the differences in elution efficiency, the affinities of phage-displayed peptides selected from either strategy were relatively similar. Although both selections yielded peptides with significantly improved affinity in comparison to KCCYSL, the improvements were modest, most likely because the parental peptide binding cannot be improved by additional amino acids.
authors.affiliation: Research Service, Harry S. Truman Veterans Memorial Hospital, Columbia, MO, 65211, USA ; Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, MO, 65211, USA.
authors.forename: Benjamin M
authors.initials: BM
authors.lastname: Larimer
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, MO, 65211, USA.
authors.forename: Jeanne M
authors.initials: JM
authors.lastname: Quinn
authors.affiliation: Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, MO, 65211, USA.
authors.forename: Kevin
authors.initials: K
authors.lastname: Kramer
authors.affiliation: Department of Biochemistry, University of Texas Health Care Center Tyler, Tyler, TX, 75708, USA.
authors.forename: Andrey
authors.initials: A
authors.lastname: Komissarov
authors.affiliation: Research Service, Harry S. Truman Veterans Memorial Hospital, Columbia, MO, 65211, USA ; Department of Biochemistry, University of Missouri, 117 Schweitzer Hall, Columbia, MO, 65211, USA.
authors.forename: Susan L
authors.initials: SL
authors.lastname: Deutscher
keywords: Affinity Maturation; ERBB2; Peptides; Phage Display