Orthogonal Peptide Purification


by Dominik Sarma

Introduction to PEC-grade peptides.

Peptide purity is often critically affected by co-eluting side products that are hard to remove by chromatographic methods. These impurities are problematic for the manufacturing of peptide APIs and lead to false-positive results during R&D activities. The latter becomes especially relevant when the time for single synthesis optimizations, such as peptide libraries, is not given.  Belyntic’s PEC-Linker provides you with a simple and straightforward tool to selectively catch your desired product from your crude mixture to reach a new dimension of purity. PEC is based on catch-and-release principles to chemo-selectively purify your peptide of interest - we call this a "PEC-grade" peptide.  Let`s start with our animation video that shows you how the principles work. 

PEC-grade is truly orthogonal.

In contrast to chromatography, the PEC-Linker chemo-selectively isolates the target peptide from its crude mixture. It, therefore, reflects a truly orthogonal approach to HPLC. The purification of the hydrophilic peptide Histone H3 (1-20), synthesized via SPPS, is a formidable challenge due to the co-eluting impurities. In collaboration with Bachem UK, we showed how to achieve high purities using the PEC-Linker. The chromatogram shown in Figure 1 highlights very nicely how impurities can hide beneath an otherwise clean-looking main peak. Only mass analysis reveals the accumulation of truncations, which all represent a potential threat to your assay. PEC efficiently removes these impurities regardless of their similarity to the target peptide. The PEC-grade Histone peptide is now ready for application. 

Figure 1: Chromatogram of Histone peptide before and after PEC purification 

Check the full Case Study "Orthogonality"

PEC-grade peptide libraries for immunological applications.

The PEC technology allows the processing of peptide purification in a 96 well-plate format, enabling easy access to purified peptide libraries. In this particular case, PEC's strength becomes apparent since high-quality peptides can be obtained in unprecedented times and at low cost. Since most peptidic impurities that may cause false-positive signals are removed, subsequent screening or validation assays' reliability is greatly enhanced. These PEC-grade peptide libraries show excellent performance in, for example, validation studies of SARS-CoV-2 epitopes, likely to elicit a T cell response. A study in collaboration with Garbi Lab from IEI Bonn highlights the parallel purification's potential during the development of in-silico designed peptide drugs or vaccines. Figure 2 shows the T-cell response of a control PEC-grade peptide with a peptide obtained from a competitor. 

Figure 2: Comparison of PEC-grade peptides with a competitor in T-cell activation assays. 

Check the full Case Study "Peptides vs Virus"

Get access to PEC-grade peptides with Belyntic's purification kits.

Belyntic offers you the first broadly applicable peptide purification kits to take advantage of PEC-grade peptides in your own research projects or manufacturing processes (Table 1). The kit comes together with related consumables and provides you with quick access to the technology.

Table 1: Overview of purification kits available in our store.

Our Research Kit works as the perfect introduction to the world of PEC peptide purification. The Ready-to-Use package comes with PEC-Linker, cartridges pre-filled with activated filter material, and related consumables. 

Download the product flyer.

We designed the Developer Kit to let pu­rification be integrated flexibly in various workflows and projects. Throughput and scale are adjustable. The kit comes with the essential materials (PEC-Linker, activated filter material) along with a manual.

Download the product flyer.

Our High-Throughput Kits open doors for the massive parallelization of peptide manufacturing. Create purified peptide libraries in the preparative scale using catch-and-release purification methodologies in 48- or 96-well filter plates. 

Download the product flyer.

Get ready for the changing tides and explore our website for more information or get in contact with us. 

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