How to proceed with identification after peptide synthesis? Bioinformatics methods have been used to predict the function of a certain sequence, and I want to verify it further.

The following aspects can be referenced for subsequent validation:

1. Peptide Identification:

• Mass Spectrometry (MS): Used to determine the molecular weight of peptides and confirm that the synthesized peptide is consistent with expectations.
• Amino Acid Sequence Analysis: Analyze the amino acid sequence of the peptide through tandem mass spectrometry (MS/MS) to ensure the sequence is correct.
• Liquid Chromatography (HPLC): Used to assess the purity of the peptide

2. Peptide Function Validation:

1) In vitro experiments: Conduct functional experiments using cell culture, such as cell proliferation, apoptosis, and pathway activation, to verify the bioactivity of the peptide.

• Enzyme Activity Test: If the peptide has potential enzyme inhibition or activation effects, its activity can be evaluated through corresponding substrate conversion experiments.
• Binding Experiments: If the predicted function involves binding to other proteins or DNA/RNA, experiments such as co-immunoprecipitation, yeast two-hybrid, or EMSA can be used to verify the peptide's binding ability to target molecules.

2) In vivo experiments (optional): If the peptide has potential biological activity, further validation of its function can be conducted in animal models. This includes evaluating pharmacokinetics (absorption, distribution, metabolism, excretion) and pharmacodynamics (dose-response relationship, efficacy, side effects).

BiotechPack, A Biopharmaceutical Characterization and Multi-Omics Mass Spectrometry (MS) Services Provider

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