Membrane Permeability of Temporin L and NAP Peptides
Disciplines
Analytical Chemistry | Medicinal-Pharmaceutical Chemistry
Abstract (300 words maximum)
Peptide therapeutics are expanding immensely due to their ability to reach target proteins, high specificity, low cost, and minimal toxicity. Today, there are over 80 therapeutics approved, and 150 that are in clinical trials seeking approval to treat a wide range of diseases and conditions. However, one of the inherent disadvantages of peptide is they have poor membrane permeability. This research aims to test the membrane permeability of some well-known peptides employing liquid chromatography and mass spectrometry. Two widely used peptides such as Temporin L and NAP were synthesized using solid phase peptide synthesis using rink-amide resin, cleaved with high concentration of trifluoracetic acid and precipitated with cold ether. After lyophilization of these peptides, they were characterized by LCMS to confirm their purity and identity. Two calibration curves were generated from two peptides varying their concentrations and obtaining their abundance using LCMS. To better validate the methods, the limit of detection (LOD) and the limit of quantification (LOQ) were calculated. Both peptides showed high sensitivity with the LOD/LOQ values being smaller than 50 picomolar. Membrane permeability was tested using Octanol-Buffer system. Peptide was mixed in the buffer solution, and octanol was added to the system and rotated for about an hour. After centrifugation, the peptide was collected from water layer and obtained their abundance using LCMS. The preliminary results showed that around 90% of Temporin L was transferred from the buffer into the octanol phase.
Use of AI Disclaimer
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Academic department under which the project should be listed
CSM – Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad Halim
Membrane Permeability of Temporin L and NAP Peptides
Peptide therapeutics are expanding immensely due to their ability to reach target proteins, high specificity, low cost, and minimal toxicity. Today, there are over 80 therapeutics approved, and 150 that are in clinical trials seeking approval to treat a wide range of diseases and conditions. However, one of the inherent disadvantages of peptide is they have poor membrane permeability. This research aims to test the membrane permeability of some well-known peptides employing liquid chromatography and mass spectrometry. Two widely used peptides such as Temporin L and NAP were synthesized using solid phase peptide synthesis using rink-amide resin, cleaved with high concentration of trifluoracetic acid and precipitated with cold ether. After lyophilization of these peptides, they were characterized by LCMS to confirm their purity and identity. Two calibration curves were generated from two peptides varying their concentrations and obtaining their abundance using LCMS. To better validate the methods, the limit of detection (LOD) and the limit of quantification (LOQ) were calculated. Both peptides showed high sensitivity with the LOD/LOQ values being smaller than 50 picomolar. Membrane permeability was tested using Octanol-Buffer system. Peptide was mixed in the buffer solution, and octanol was added to the system and rotated for about an hour. After centrifugation, the peptide was collected from water layer and obtained their abundance using LCMS. The preliminary results showed that around 90% of Temporin L was transferred from the buffer into the octanol phase.