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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

     


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