Peptidomics is the study of endogenous polypeptides and small molecular proteins in organisms, especially in blood, cerebrospinal fluid and body fluids. These endogenous polypeptides have many signal regulating molecules, such as cytokines, growth hormone, hormone peptide and so on which can provide a lot of valuable information. There are also many small molecule proteins and peptides. Some of their functions are unknown, and some belong to disease specific degradation fragments of some proteins in circulating fluid. These soluble fragments can provide abundant information about disease status, efficacy, toxicity and so on.
Advantages:
No antibodies: antibodies have less effective on peptides that are less than 10kd;
Accurate identification of peptide sequences: peptides are ideal peptide biomarkers. Based on mass spectrometry, peptides can be identified accurately and repeatedly;
Efficient enrichment efficiency: more than 99% of the protein mass is removed in the sample treatment;
Rapid construction method.
Application areas:
Biomarker discovery / validation: identification and validation of potential disease markers / drug targets;
Side effects: detection of nonspecific and unwanted drug effects, such as degradation;
Sample quality control: peptide segments, as degradation products, can indicate the quality of the sample.
Examples:
Objective
Identify polypeptides in human milk to study their functions.
Sample
5 mothers’ milk for twenty-eighth days
Experimental procedure
A. Milk samples: remove the lipid and protein, and purify the polypeptide of the sample;
B. LC-MS/MS mass spectrometry analysis: exclustion list increased the number of identification;
C. Data analysis: polypeptide identification and endogenous fracture location analysis;
D. Functional experiments: antibacterial experiments of purified peptides from milk.
Experimental results
In milk samples, more than 300 peptides of 37 proteins were β-casein. But other milk proteins such as lactoferrin, lactalbumin and immunoglobulin A were not identified. By comparing the identified peptides to their corresponding protein sequences, it was found that endogenous polypeptides only came from certain proteins and protein specific sequences. Many identified milk peptides were similar to known antimicrobial peptides or immunomodulatory peptide sequences. Milk purified peptides had also been shown to inhibit the growth of Escherichia coli and Staphylococcus aureus.
Creative Proteomics provides custom tailed peptidomics service for your research. For more information, please feel free to contact us.
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Website: https://www.creative-proteomics.com