Welcome to the Biotage Peptide Synthesis Blogs.

      Optimizing the removal of an STmp protecting group

      Dec 29, 2020 3:29:00 PM / by Elizabeth Denton posted in Chromatography Fundamentals, Peptides, Troubleshooting and Optimization

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      Disulfide rich peptides are unique in both their incredibly high cysteine content, but also in the stability imbued by the multiple disulfide bonds.  These peptides, stable under extreme conditions that would either denature or degrade a similar linear peptide, make disulfide rich peptides attractive as both therapeutics or as scaffolds upon which to construct non-native functionality.  Synthesizing these compounds, however, still remains a challenge.

      I have discussed previously strategies that enable on-resin chemistry via orthogonal protecting groups.  These groups can be removed under mildly acidic, metal catalyzed, or even oxidizing conditions.  In today’s post, I’ll demonstrate the utility of using disulfide shuffling as a cysteine protection strategy.

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      Green solvents for solid phase peptide synthesis

      Dec 21, 2020 3:29:00 PM / by Elizabeth Denton posted in Chromatography Fundamentals, Synthesis

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      As peptide therapeutics continue to gain interest from the medical community and pharmaceutical companies, concerns regarding the cost of manufacturing also grow.  Cost  includes the expense of reagents and solvents, including DMF and NMP, used in the synthesis but also subsequent disposal.  

      With growing motivations to improve the "green" characteristics of chemistry (reducing waste, reducing use of harmful solvents, etc), today's post will highlight some recent work evaluating alternative solvents for use in solid phase peptide synthesis.

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      What mobile phase flow rate should I use for my peptide purification with flash chromatography?

      Dec 17, 2020 3:28:00 PM / by Elizabeth Denton posted in Chromatography Fundamentals, Peptides, Reversed-phase

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      I’ve recently worked with several peptide groups that are trying out flash purification with their peptides for the first time.  And it never fails, every single interaction includes the question “what flow rate should I use for these cartridges?”

      There is a lot of information available highlighting optimal flow rates for HPLC method development, but very little information for larger particle stationary phases.  I personally have used a wide range of flow rates for my peptide purification with differing outcomes.  So in today’s post I’d like to have a more thorough discussion about mobile phase flow rate and it’s impact on your chromatography.

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      How to purify peptides using a step gradient in flash column chromatography

      Dec 8, 2020 3:27:00 PM / by Elizabeth Denton posted in Chromatography Fundamentals, Peptides, Reversed-phase

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      Flash chromatography can be a challenging technique for peptide purification due to the lower resolution achieved with large particles.  While some may see this as a disadvantage, the significantly greater loading capacity gives me reason to make this work. So how can I achieve the high purity levels often accessed using traditional reversed-phase HPLC methods?

      In this post, I’ll discuss using a step gradient for peptide purification.  Step gradients are commonly used in normal-phase small molecule purification and typically improve the purification efficiency while reducing the overall purification time.

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      Optimizing a mobile phase gradient for peptide purification using flash column chromatography

      Dec 3, 2020 3:27:00 PM / by Elizabeth Denton posted in Chromatography Fundamentals, Peptides, Reversed-phase, Workflow

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      Have you ever wondered if there was a faster (and possibly cheaper) way to purify your peptides?

      My colleagues and I in the peptide community rely almost exclusively on reversed-phase HPLC for delivery of highly pure peptide products.  However, this process is often very time consuming and requires expensive columns and solvents to be successful.  Alternatively, peptide purification via reversed-phase flash column chromatography can be used to complete a purification in a fraction of the time and with a fraction of the costs.

      Here I will show how I do gradient optimization for peptide purification via reversed-phase flash column chromatography and will highlight the similarities with standard HPLC methodologies.

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