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

@article{Hilpert2007PeptideAO,
  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},
  year={2007},
  volume={2},
  pages={1333-1349}
}
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… 
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References

SHOWING 1-10 OF 46 REFERENCES
Spot synthesis: observations and optimizations.
TLDR
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
TLDR
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
TLDR
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.
TLDR
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
TLDR
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.
TLDR
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
TLDR
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.
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