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Analytical Method Development and Validation
Peptides are widely found in nature and have important physiological effects on the human body. In recent years, with the vigorous development of life science research, the research progress on peptides is changing with each passing day. Peptides have shown magical research and application value in immune function, information transmission, cell secretion, precursor signal, disease occurrence and treatment.
Introduction to Detection Methods
Currently, high performance liquid chromatography (HPLC), capillary electrophoresis (CE), mass spectrometry (MS) and their associated technology have become a primary means of measuring the peptide, and are being more widely used.
High performance liquid chromatography (HPLC)
HPLC is a new separation and analysis technology developed on the basis of classical liquid chromatography and gas chromatography. It has the characteristics of high separation efficiency, high speed, easy operation, good detection sensitivity and wide application range to samples. It has been widely used for the analysis and determination of peptides. Reversed-phase high performance liquid chromatography (RP-HPLC) has a very high resolution for the analysis and purification of non-polar small molecule peptides with a molecular mass below 1000.
Capillary electrophoresis (CE)
CE is a kind of liquid phase separation technology that uses a high-voltage electric field as a driving force and a capillary as a separation channel to separate components according to the difference in mobility and distribution between components in a sample.
Mass spectrometry (MS)
The mass spectrum of a peptide detected by mass spectrometry (MS) is usually referred to as a fingerprint. The biggest advantage of mass spectrometry is stability, reproducibility and high accuracy. At present, mass spectrometry is applied to peptide detection mainly by matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS), liquid chromatography-mass spectrometry (LC-MS) and capillary electrophoresis-mass spectrometry (CE-MS).
Hydrophobic interaction chromatography (HIC)
HIC is the use of a hydrophobic group in a peptide to create a hydrophobic interaction with a stationary phase for separation analysis. Iron-Exchange chromatography (IEXC) can be used to isolate and purify biologically active peptides under neutral conditions using different chargeability of the peptide. Methods for detection of peptide analysis also include chromatography of membrane protein (CMP), high-performance displacement chromatography (HPDC), perfusion chromatography (PC), affinity chromatography (AC).
In addition to the above methods, amino acid composition analysis, amino acid sequence analysis, field analytical mass spectrometry, IR, UV spectroscopy, CD, circular dichroism, bioassay, radioisotope labeling and immunological methods have been applied to analyze the peptides characterization.
- Extraction of peptides: protease inactivation
- Pretreatment: removal of protein
- Peptide purification: chromatography, capillary gel
- Peptide identification: MALDI-TOF-MS, LC-MS, CE-MS
- Peptide quantification: immunoassay, isotope mass shift
By removing all proteins in biological samples, body fluids, and tissue extracts by size exclusion chromatography, LC-MS-based assays can be quickly established and validated to identify peptides with high precision and repeatability. After a detailed and comprehensive discussions with customers, we will provide you with a peptide set research and analysis services tailored.
Creative Peptides has successfully developed a reliable method for the analysis of peptides by using several of the peptide analysis methods mentioned above. Creative Peptides is dedicated to the detection and analysis of peptides and provides a powerful tool for research in the fields of protein chemistry such as medicine, clinical pathology and preventives. Our experienced scientists will tailor the most appropriate method and 100% guaranteed service to our customers. Please contact us.
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