Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion

  title={Peptide arrays on cellulose support: SPOT synthesis, a time and cost efficient method for synthesis of large numbers of peptides in a parallel and addressable fashion},
  author={Kai Hilpert and Dirk F. H. Winkler and Robert E. W. Hancock},
  journal={Nature Protocols},
Peptide synthesis on cellulose using SPOT technology allows the parallel synthesis of large numbers of addressable peptides in small amounts. In addition, the cost per peptide is less than 1% of peptides synthesized conventionally on resin. The SPOT method follows standard fluorenyl-methoxy-carbonyl chemistry on conventional cellulose sheets, and can utilize more than 600 different building blocks. The procedure involves three phases: preparation of the cellulose membrane, stepwise coupling of… 
The spot technique: synthesis and screening of peptide macroarrays on cellulose membranes.
This chapter presents the basic procedures and screening methods related to spot synthesis and outline protocols for easy-to-use detection methods on these peptide arrays.
SPOT Synthesis of Peptide Arrays on Self‐Assembled Monolayers and their Evaluation as Enzyme Substrates
By combining the robustness and versatility of SAM-coated gold platforms and the easy-to-handle SPOT peptide synthesis methodology, an efficient method for preparing peptide arrays on gold surfaces is described herein.
Synthesis of peptide arrays using SPOT-technology and the CelluSpots-method.
This method allows the synthesis of low-cost peptide arrays containing around 900 large spots of addressable peptides on a cellulose sheet of 19 cm x 29 cm.
Peptide arrays with a chip.
This paper presents a combinatorial synthesis based on electrically charged solid amino acid particles that should allow for the translation of entire genomes into a set of overlapping peptides to be used in proteome research.
Continuous-flow solid-phase peptide synthesis: a revolutionary reduction of the amino acid excess.
A highly efficient continuous-flow technique for the synthesis of peptides was developed and exotic and expensive artificial amino acids were incorporated into peptidic sequences by the utilization of a reasonable number of amino acid equivalents.
Flow-through synthesis on Teflon-patterned paper to produce peptide arrays for cell-based assays.
The confinement and flow-through mixing significantly improves the peptide yield and simplifies the automation of this synthesis, which was used in cell-based screening to identify 14 potent bioactive peptides that support the adhesion or proliferation of breast cancer cells in a 3D environment.
Peptide Arrays for Binding Studies of E3 Ubiquitin Ligases.
The implementation of SPOT binding assays with focus on the identification of N-recognin substrates, applicable also for plant NERD enzymes are described.
Heat-enhanced peptide synthesis on Teflon-patterned paper.
It is demonstrated that IR-heating in solid phase peptide synthesis shortened the reaction time necessary for amide bond formation down to 3 minutes; in some couplings of alpha amino acids, conversion rates increased up to fifteen folds.
SPOT synthesis as a tool to study protein-protein interactions.
Protocols for the peptides synthesis on cellulose including the preparation of different cellulose membranes and easy-to-use detection methods on these peptide macroarrays are presented.
Peptide library synthesis on spectrally encoded beads for multiplexed protein/peptide bioassays
A new technology for measuring antibody-peptide interactions in vitro that leverages spectrally encoded beads for biological multiplexing and is broadly applicable to any range of peptide screening applications, with the capability to multiplex into libraries of hundreds to thousands of peptides in a single assay.


Spot synthesis: observations and optimizations.
The most important results were: the signal intensity of ligate binding to cellulose-bound peptides and the affinity of the corresponding soluble peptides show good correlation, and optimization of side-chain deprotection revealed that a 30-min pretreatment of the cellulose with 90% trifluoroacetic acid followed by the standard deprotected protocol resulted in higher purity of the synthesized products.
General method for the rapid solid-phase synthesis of large numbers of peptides: specificity of antigen-antibody interaction at the level of individual amino acids.
  • R. Houghten
  • Biology, Chemistry
    Proceedings of the National Academy of Sciences of the United States of America
  • 1985
Through examination of the binding of these analogs to monoclonal antibodies raised against residues 75-110 of HA1, it was found that a single amino acid, aspartic acid at position 101, is of unique importance to the interaction.
Individually addressable parallel peptide synthesis on microchips
An efficient and flexible method for the parallel synthesis of peptides on individually addressable microchips, using digital photolithography and photogenerated acid in the deprotection step, allowing potential development of peptide microch chips for various pharmaceutical and proteomic applications in routine research laboratories.
Improved performances of spot multiple peptide synthesis.
A new software for the design of peptides to be prepared by the manual Spot synthesis method is developed and most of the common protocols for epitope mapping are covered and it is shown that peptides of up to 29 residues were immunoreactive.
Automated Synthesis of Solid-Phase Bound Peptides
The SPOT method was developed by Ronald Frank for simultaneous multiple peptide synthesis on separate sites on a homogeneous membrane carrier (Frank 1992). The principle of the technique is to
Cellulose-bound Peptide Arrays: Preparation and Applications
SPOT synthesis was developed by Ronald Frank and co-workers and first presented the method in 1990 at the 21st European Peptide Symposium.
Characterizing and optimizing protease/peptide inhibitor interactions, a new application for spot synthesis.
New peptides were designed in a step-wise fashion which not only inhibited elastase 400 times more strongly than the original peptide, but are highly specific for the enzyme.
Use of luminescent bacteria for rapid screening and characterization of short cationic antimicrobial peptides synthesized on cellulose using peptide array technology
A protocol for screening large numbers of peptides against any microbe of interest and demonstrates good correlation with conventional killing or minimal inhibitory concentration assays performed with the same peptides synthesized by standard solid-phase peptide synthesis.