AI Article Synopsis
- The SPOT technology allows for efficient synthesis of soluble peptides in the microgram range, specifically utilizing new acidic labile linkers HMPA and HMPB for improved outcomes.
- This method minimizes side-chain reactions that typically occur during traditional alkaline peptide cleavage, resulting in higher purity peptides.
- The automated process is time- and cost-effective, making it suitable for producing a large number of peptides for various biological applications like T-cell epitope screening and vaccine development.
Article Abstract
The SPOT technology can fulfill most requirements for highly parallel, multiple peptide synthesis of soluble peptides within the upper microgram range. Here, we report on an improved method using hydroxymethylphenoxyacetic acid (HMPA) for 19 amino acids and 4-(4-hydroxymethyl-3-methoxyphenoxy)-butyric acid (HMPB) for proline as acidic labile linkers in SPOT synthesis. Using this approach we could reduce side-chain reactions normally occurring during conventional alkaline peptide cleavage from cellulose membranes. All synthesis steps were adapted to fully-automated SPOT synthesis and therefore represent a time- and cost-saving procedure. Furthermore, the improved cleavage and washing steps resulted in peptides with authentic C-termini in a purity range of 60-95%. Our improved method is ideal for synthesizing many thousand different peptides subsequently used directly for different biological assays requiring authentic C-termini, such as CD8 T-cell epitope screening, vaccine immunization, or tumor imaging.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.bmcl.2008.05.116 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Drosophila X virus-like particles as delivery carriers for improved oral insecticidal efficacy of scorpion Androctonus australis peptide against the invasive fruit fly, Drosophila suzukii.
Insect Sci
June 2024
Faculty of Bioscience Engineering, Department of Plants and Crops, Ghent University, Ghent, Belgium.
Insect-specific neurotoxic peptides derived from the venoms of scorpions and spiders can cause acute paralysis and death when injected into insects, offering a promising insecticidal component for insect pest control. However, effective delivery systems are required to help neurotoxic peptides pass through the gut barrier into the hemolymph, where they can act. Here, we investigated the potential of a novel nanocarrier, Drosophila X virus-like particle (DXV-VLP), for delivering a neurotoxin from the scorpion Androctonus australis Hector (AaIT) against the invasive pest fruit fly, Drosophila suzukii.
View Article and Find Full Text PDFDissecting the Structural Dynamics of Authentic Cholesteryl Ester Transfer Protein for the Discovery of Potential Lead Compounds: A Theoretical Study.
Int J Mol Sci
July 2023
MOE Key Laboratory for Nonequilibrium Synthesis and Modulation of Condensed Matter, School of Physics, Xi'an Jiaotong University, Xi'an 710049, China.
Current structural and functional investigations of cholesteryl ester transfer protein (CETP) inhibitor design are nearly entirely based on a fully active mutation (CETP) constructed for protein crystallization, limiting the study of the dynamic structural features of authentic CETP involved in lipid transport under physiological conditions. In this study, we conducted comprehensive molecular dynamics (MD) simulations of both authentic CETP (CETP) and CETP. Considering the structural differences between the N- and C-terminal domains of CETP and CETP, and their crucial roles in lipid transfer, we identified the two domains as binding pockets of the ligands for virtual screening to discover potential lead compounds targeting CETP.
View Article and Find Full Text PDFCapsids of hepatitis B virus e antigen with authentic C termini are stabilized by electrostatic interactions.
FEBS Lett
March 2020
Protein Expression Laboratory, NIAMS, National Institutes of Health, Bethesda, MD, USA.
The hepatitis B virus e antigen, an alternative transcript of the core gene, is a secreted protein that maintains viral persistence. The physiological form has extended C termini relative to Cp(-10)149, the construct used in many studies. To examine the role of the C termini, we expressed the constructs Cp(-10)151 and Cp(-10)154, which have additional arginine residues.
View Article and Find Full Text PDFPeriplasmic Nanobody-APEX2 Fusions Enable Facile Visualization of Ebola, Marburg, and Mĕnglà virus Nucleoproteins, Alluding to Similar Antigenic Landscapes among and .
Viruses
April 2019
Disease Intervention and Prevention, Texas Biomedical Research Institute, 8715 W. Military Dr., San Antonio, TX 78227-5302, USA.
Article Synopsis
- This study examines the use of evolved soybean ascorbate peroxidase (APEX2) as a reporter by fusing it to llama nanobodies (single-domain antibodies) to enhance periplasmic expression, which maintains the natural structure of antibodies.
- The researchers successfully purified significant amounts of these fusion proteins and utilized them in various detection methods like Western blotting and ELISA to visualize viral nucleoproteins.
- The findings suggest that these sdAb-APEX2 fusions can improve detection of diverse viruses, including a newly discovered virus, and facilitate ongoing studies of various Filovirus species in a controlled laboratory environment.
Production and purification of recombinant human hepcidin-25 with authentic N and C-termini.
J Biotechnol
February 2015
Univ Bordeaux, BPRVS, EA 4135, F-33000 Bordeaux, France; Bordeaux INP, BPRVS, EA4135, F-33000 Bordeaux, France.
Hepcidin was first identified as an antimicrobial peptide present in human serum and urine. It was later demonstrated that hepcidin is the long-sought hormone that regulates iron homeostasis in mammals. Recombinant human Hepcidin-25 (Hepc25) was expressed in Pichia pastoris using a modified version of the pPICZαA vector.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!