In my role as a peptide application scientist, I have had the pleasure of working with many groups that are venturing into the world of peptides for the first time.  Although it seems rather  straightforward to experienced synthetic chemists, producing acceptable yield and purity certainly comes with unique challenges in solid phase peptide synthesis .

In this post I would like to present some of the tips and tricks that I have picked up along the way.

More and more groups are exploring the utility of peptides with an ever widening variety of applications. And although peptides are getting cheaper to purchase outright, many groups are continuing to bring peptide synthesis in house. As more groups join the peptide community, I frequently encounter questions about the basics of peptide synthesis.

In today's post, I'd like to cover a little history of solid phase synthesis as well as highlight some differences between the chemistries.

Since the development of Fmoc-based solid phase peptide synthesis, a wide variety of cleavage cocktails have emerged.  Each cleavage cocktail contains a unique combination of scavengers designed to prevent either side reactions mediated by the released protecting groups or the side chains themselves, or both during the peptide cleavage reaction.  As the number of scientists performing peptide synthesis grows, the question “which cleavage cocktail should I use?” comes up more often than not.

In today’s post, I’ll highlight the role of of scavengers for peptides containing cysteine residues.

Chemical synthesis of peptides, and even proteins, offers the possibility to expand the functionality and stability imbued by nature.  However, chemical synthesis of very long peptides and small proteins remains today an exceedingly difficult task.  Several ligation strategies have been developed that help to alleviate this challenge.  These strategies though, require a purified, yet fully protected peptide fragment.

Purification of a fully protected peptide species can be challenging by standard reversed-phase techniques, primarily due to the limited solubility of protected peptides in aqueous solutions.  In today’s post, I will discuss using normal-phase chromatography for purification of protected peptides.

Orthogonal amino acid protecting groups effectively expand the chemical tool kit available to peptide chemists allowing for synthesis of much more complex molecules.  Often times, orthogonal protecting groups are used in Fmoc-based chemistry to facilitate post-synthesis modifications of peptides, like the addition of small molecule fluorophores and more commonly now, peptide cyclization efforts.

In a previous post, I discussed optimizing the removal of an ivDde protecting group.  In today’s post, I’ll explore the removal of an alloc protecting group from a lysine residue.

What is the main goal of a peptide chemist? Elizabeth Denton, Ph.D., explains how Biotage sees the peptide synthesis workflow and how we focus on shortening the process time for scientists.

 It never fails. A sequence looks relatively straightforward on paper and it's of moderate length so you start synthesizing without too much thought into the protocol. Your synthesis is finished and you run the analytical HPLC but your product is nowhere to be found (or only very small quantities). So what went wrong? And what can you do differently to increase your crude yield and purity?

Elizabeth Denton, PhD, Senior Scientist Peptide Chemistry, recorded a webinar titled Disulfide Rich Peptides: Optimizing and Automating Syntheses with Regioselective Formation of Disulfide Bonds. To learn more, read the description below as well as watch the recording!