Analyte Peptidomimetics Selected from Phage Display Peptide Libraries: A Systematic Strategy for the Development of Environmental Immunoassays

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• 作者：Soledad Cardozo ; André ; s Gonzá ; lez-Techera ; Jerold A. Last ; Bruce D.Hammock ; Karl Kramer ; and Gualberto G. Gonzá ; lez-Sapienza
• 刊名：Environmental Science & Technology
• 出版年：2005
• 出版时间：June 1, 2005
• 年：2005
• 卷：39
• 期：11
• 页码：4234 - 4241
• 全文大小：182K
• 年卷期：v.39,no.11(June 1, 2005)
• ISSN：1520-5851

文摘

Due to their simplicity, speed, low cost, and specificity,immunoassays have become a useful tool for the analysisof environmental pollutants. Once the anti-haptenantibodies are produced, the same hapten or a relatedmolecule is conjugated to a tracer enzyme or coating proteinto set up the assay. Here we report the use of peptidesthat mimic the analyte as advantageous substitutes ofcompeting haptens. These peptides, which open opportunitiesfor innovation in the development of tracer reagents,can be selected from phage display libraries in astraightforward systematic manner. The concept wasproven using assays for the herbicides molinate and atrazineas model systems. Several characteristics of the selectionprocess that may affect the final assay were analyzed,such as the phage coat proteins fused to the peptide, theuse of linear or constrained peptide libraries, the effectof the concentration of analyte used during the selectionprocess, and the use of monoclonal or polyclonal antibodiesas selector molecules. In all cases we found that theselected peptides performed with improved sensitivity ascompared with the chemical hapten conventional assays,showing an analogous cross-reactivity pattern. Interestingly,the phage particles perform as robust and highlystandardized assay reagents, and due to their filamentousrepetitive structure, they function as sensitive multienzymaticreporters.