Hits:1856 , posted at 2023/11/15 10:41
Using a CSBio research scale peptide synthesizer, we share a case study
of how a 132-mer peptide can be synthesized with high purity
Introduction
Synthesis of peptides at any length requires
extreme attention to detail for accuracy and purity of the desired compound.
Correctly synthesizing peptides of 50 mer length is difficult, over 100 mer
length is a remarkable feat. CSBio’s fully automated instruments are
purposefully designed to maximize resin and solvent interaction while
maintaining a clean system throughout the synthesis protocol, making it
possible to consistently synthesize peptides containing over 130 amino acids at
very high purities.
Brief
The chemists in CSBio’s in-house cGMP
peptide manufacturing facility were tasked with making a cyclic peptide 132
amino acids in length containing a Cysteine to Cysteine disulfide bond linking
the 39th and 79th amino acids.
132
AA peptide sequence (protected)
Using a research scale CSBio peptide synthesizer, Fmoc
protected amino acids were attached one by one in a stepwise fashion.
Engineering decisions such as 180° inversion mixing as well as dedicated lines
to separate the delivery of amino acids and piperidine ensure more resin to
solvent interaction than nitrogen bubbling, vortex or oscillation mixing, while
also maximizing coupling with no risk of chemical cross contamination.
CSBio
CS136X
Upon
peptide synthesis completion the resin was cleaved giving the chemist’s 620mg
of crude at 75% purity. A portion of the crude (220mg) was loaded into a 1” C18
HPLC column, cyclization of the fractions was performed using an iodine.
Post purification the chemists were able to extract 18mg of the
re-purified disulfide peptide which tested at 98.7% purity. Resulting in an
8.2% yield from crude for a compound whose molecular weight was >15000g/mol.
132mer
post-purification chromatogram
Conclusion
CSBio synthesizers are designed to deliver the user with the highest
crude quality for any possible peptide. As the only automated synthesizer
manufacturer that produces cGMP peptides, constant collaboration of in-house
chemists and engineers during the design process enables CSBio to improve
synthesis results compared to industry competitors; making CSBio synthesizers
the most robust and reliable instruments on the market for synthesis of any
peptide.
Interested in our research scale peptide synthesizers? Get in touch with us.
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