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Peptide Synthesis Application and Recommended Purity

Custom synthetic peptides are valuable reagents in biological research, and are widely used to mimic complex protein structures or antigens. Quality of custom peptides, i.e., purity, is the key to success and integrity of your research. Consequently, the main cost for peptide synthesis is incurred on purification and characterization of custom peptides. We have enlisted some of common applications of synthetic peptides and recommended purity for custom synthesis based on our consultations with experts familiar with these biological assays.

Research area	Typical assay	Application	Recommended quality
Antibody . antigen interactions	ELISA, Fluorescence polarization assay, Fluorescence resonance energy transfer assay (FRET), Surface Plasmon Resonance (SPR)	1. to define minimal epitope for Ab binding 2. to substitute or mimic complex protein or polysaccharide antigens	> 80% purity for ELISA in which peptides are coated in excess on solid plate surface > 90% purity for in solution assays and SPR assays
Antigenic peptides in immune lymphocyte assays	⁵¹Cr release assay, ³H thymidine incorporation assays, T cell cloning and culturing, T cell cytokine production assays	Used as antigens in culture supernatant to sensitize target cells or antigen presenting cells for T cell recognitions	> 80% purity
Antigenic peptides in quantitative T cell assays	ELISPOT assay, intracellular cytokine staining assay	1. Commonly used as overlapping peptide pools to represent open reading frame of antigens. 2. Truncated peptide series is useful to map T cell epitope	> 90% purity for common research assays > 95% for clinical or validated assays
Peptide conjugate vaccines	Vaccination to generate peptide specific immune sera as research reagents for Western blot or immunoprecipitation	Commonly conjugated to KLH or other protein carrier to immunize animals for peptide-specific antibodies	> 95% purity for optimal conjugation efficiency
MHC Tetramer complex	Tetramer analysis of epitope-specific T cells in flow cytometry	Construction of epitope specific MHC tetramer complex as flow cytometry reagents	> 95% purity for optimal tetramer folding efficiency
Structural studies	NMR, crystallization		> 98% purity for optimal interpretation of structural information
Enzymatic substrates	In vitro assays to assess kinases, phosphatases, or proteases activities	Commonly used as substrates in these enzymatic assays. Can be assessed with fluorescent or other chemical labels	> 90% for research and high throughput screening assays > 95% purity for quantitative analysis
Biological substrates for metabolic or anti-microbial activities, or cell penetrating peptides for gene or drug delivery	Animal experiments involving peptides, cell culture assays with peptides, transfection, gene delivery	1. Small peptides have biological functions, such as defensin peptides, or can act as inhibitors to block biological functions, such as T-20 peptide of HIV gp41 to inhibit viral entry. These peptides are studied as peptide drugs in animal models 2. A class of small peptides has been identified for their ability to permeate cell membrane and thus explored as delivery tools for gene, siRNA or drug	> 95% purity for in vitro studies > 98% for animal studies