Multi-pin peptide synthesis strategy for T cell determinant analysis.
Techniques to synthesize many peptides simultaneously exist, however their individual cleavage and subsequent purification constitutes a bottleneck to total throughput. Biological screening of peptides is generally carried out at physiological pH in aqueous solutions. However, peptides, unless individually purified are usually contaminated by residual compounds used in their preparation such as trifluoroacetic acid, organic solvents, scavengers etc. In testing with cellular systems, such as T cell determinant analysis, such contaminations must be rigorously excluded. We have extended the pin synthesis technique of synthesizing and screening large number of peptides (Geysen et al., 1984) to the analysis of T cell determinants. Peptides can be synthesized on polyethylene pins, the side chain protective groups removed and the peptides washed free of contaminants. A linker system stable under these conditions can then be triggered to cleave the peptides from the pins in an aqueous solution at neutral pH. This strategy enables the rapid mapping of T cell determinants. It is also applicable to other systems where large numbers of solution phase peptides are required, for example, in the study of hormone analogues.
Maeji
N J
NJ
Coselco Mimotopes Pty. Ltd., Clayton, Victoria, Australia.
Bray
A M
AM
Geysen
H M
HM
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D054659:Diketopiperazines; D000939:Epitopes / Q000276:immunology*; D008213:Lymphocyte Activation / Q000276:immunology; D008722:Methods; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis*; D010879:Piperazines / Q000737:chemistry; D011095:Polyethylenes; D013601:T-Lymphocytes / Q000276:immunology*; D013744:Tetanus Toxin / Q000737:chemistry
D054659:Diketopiperazines; D000939:Epitopes; D010455:Peptides; D010879:Piperazines; D011095:Polyethylenes; D013744:Tetanus Toxin