Peptide-Based Drug Design

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The Spot Technique: Synthesis and Screening of Peptide
Macroarrays on Cellulose Membranes
Dirk F.H. Winkler and William D. Campbell
Summary
Peptide arrays are a widely used tool for drug development. For peptide-based drug
design it is necessary to screen a large number of peptides. However, there are often
difficulties with this approach. Most common peptide synthesis techniques are able to
simultaneously synthesize only up to a few hundred single peptides. Spot synthesis is a
positionally addressable, multiple synthesis technique offering the possibility of synthesizing
and screening up to 10,000 peptides or peptide mixtures on cellulose or other
membrane surfaces. In this chapter we present the basic procedures and screening methods
related to spot synthesis and outline protocols for easy-to-use detection methods on these
peptide arrays.
Key Words: Spot synthesis; peptide array; screening; detection; probing; HRP; POD;
biotin-labeling; peptide-protein interaction.
1. Introduction
Peptide arrays are widely used for the investigation of peptide–protein
and protein–protein interactions. Screening peptides for potentially active
compounds using peptide arrays is a very convenient tool in the development
of drug candidates. For peptide-based drug design, using peptide microarrays
it is possible to screen a high number of peptides on a small chip. Due to the
miniscule amounts of peptides synthesized directly on chips, this approach has
proven to be very difficult and often solution phase assays and analysis are
unreliable. These disadvantages can be overcome using peptide macroarrays.
The mostly widely used macroarray is produced by synthesis utilizing the spot
From: Methods in Molecular Biology, vol. 494: Peptide-Based Drug Design
Edited by: L. Otvos, DOI: 10.1007/978-1-59745-419-3 4, © Humana Press, New York, NY
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