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

• Published in: Nature protocols Vol. 2; no. 6; pp. 1333 - 1349
• Main Authors: Hilpert, Kai, Winkler, Dirk FH, Hancock, Robert EW
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 01.06.2007
• Subjects: Amino Acid Sequence; Amino acids; Automation; Cellulose; Cellulose - chemistry; Combinatorial Chemistry Techniques; Epitope Mapping - methods; Libraries; Peptide Library; Peptides; Peptides - chemical synthesis; Peptides - chemistry; Properties; Protein Array Analysis - economics; Protein Array Analysis - methods; Protocol; Synthesis; Time Factors; United States; Vancouver British Columbia Canada; Canada
• ISSN: 1754-2189; 1750-2799
• DOI: 10.1038/nprot.2007.160

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 amino acids and cleavage of the side-chain protection groups. If necessary, peptides can be cleaved from the membrane for assays performed using soluble peptides. These features make this method an excellent tool for screening large numbers of peptides for many different purposes. Potential applications range from simple binding assays, to more sophisticated enzyme assays and studies with living microbes or cells. The time required to complete the protocol depends on the number and length of the peptides. For example, 400 9-mer peptides can be synthesized within 6 days.