Peptide Purity Analysis

Peptide purity, measured by analytical reversed phase HPLC, is the percentage of the peptide compared to impurities that absorb at 210-220 nm(peptide bond absorption wavelength). Peptide purity does not take into account water and salts usually present in the sample. Sometimes alternative methods such as ion exchange, dialysis, precipitation or gel filtration chromatography are also used. Purity of peptide is a critical factor that affects the success and meaningful outcome of a research project, thus peptide purity analysis, detection of contamination and residual content are widely performed in various industries. Common contaminates of peptide product are resulted from chemical synthesis, culturing medium, or incorporated from extraction and purification procedures. Impurities consist of deletion sequences, incompletely deprotected sequences, truncated sequences and side products of the synthesis process. Residual salt and water that do not absorb at this wavelength are not quantified. In experimental planning, it is important to consider that some peptide or contamination may be toxic to cells. This kind of toxicity o cells can generally be avoided by purifying the peptide with proper methods, which will result in only trace amounts of toxic impurities.

Recommended Purity Based on Application of Peptide:

95% to 99%
Recommended for quantitative assays and structural studies.
  • Quantitative Blocking and Competitive Inhibition Assays
  • Quantitative Receptor-Ligand Interaction Studies
  • Quantitative Phosphorylation Studies & Quantitative Proteolysis Studies
  • In-vivo Studies & In-vitro Bioassays
  • Generating Standard Curves
  • Reference Standards & Diagnostic Kits
  • NMR Spectroscopy Studies & Crystallography
  • Clinical Trials & Drug Studies
  • SAR Studies
85% to 94%
Suitable for most qualitative and semi-quantitative biochemical assays.
  • Phosphorylation Studies
  • Semi-quantitative Enzyme-Substrate Studies
  • Non-quantitative Peptide Blocking Studies for Western Blotting and Immunocytochemistry
  • In-vitro Bioassays
  • Cell Attachment Studies
  • Epitope Mapping
  • Coupling to Chromatography Resins for Affinity Purifications
  • Antibody- Antigen Interaction Studies
  • Antigenic Peptides in Immune Lymphocyte Assays
70% to 84%
Suitable for a number of screening and research applications as shown right.
  • Peptide Array and Screening Studies
  • Polyclonal Antibodies
  • Immunogenic Identification
  • Mutation Screening
  • Sequence Optimization
  • ELISA Tests

Peptides with different purities (Analyzed by HPLC absorbance at 214 nm) Figure 1. Peptides with different purities (Analyzed by HPLC absorbance at 214 nm)

Creative proteomics has already developed highly sensitive, reversed phase HPLC platform to analyze peptide purity.

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