With our PEC technology in hand, we consistently create valuable knowledge about its enormous application potential - during our internal R&D activities as well as together with our partners and customers. We aim to share this information with our community to support our value propositions with a solid scientific and well-investigated basis.
Improved ecological efficiency
Chromatographic purification of peptides consumes large amounts of organic solvent. Belyntic‘s PEC technology offers remedy by drastically lowering the use of organic solvents, improving your ecological efficiency.
Results at a glance
- Improve the crude purity with DMF-free SPPS
- Reduce waste by 75% using orthogonal PEC purification
- Use the TFA catch method to enable precipitation and ether-free transfer from synthesis to purification
Green manufacturing of Bivalirudin with PEC
Due to solvent waste and energy consumption, the manufacturing of pharmaceutical ingredients has a significant environmental impact. Therefore, the industry demands new and innovative techniques for greener drug production. This study highlights DMF-free solid-phase peptide synthesis (SPPS) and its compatibility with the low-solvent consuming Peptide Easy Clean (PEC) purification technology to manufacture Bivalirudin in a green approach. We also highlight a novel TFA (Trifluoroacetic acid) catch method to eliminate the post-SPPS precipitation step, which consumes a large amount of hazardous ether.
Results at a glance
- reduce the total waste generation of single run purifications by up to 75% to 85%
- purify highly hydrophilic peptides such as histone fragments efficiently in a single step
- apply capping to get high-quality crude and final product purity
Less is more
Due to its repetitive cycles of deprotection, washing, and coupling, SPPS is a poorly atom efficient process, resulting in the consumption of a considerable amount of solvents and reagents. Furthermore, typical peptide purification with RP-HPLC also consumes a high amount of organic solvents. In this study, we compare the solvent consumption and waste generation for peptide manufacturing between RP-HPLC and the PEC purification technology.
Advanced peptide modification
In recent times, chemical modifications have opened up new perspectives for the application of peptides as drugs or conjugates in general. However, with that development come increasingly challenging peptide purification processes. Belyntic’s PEC technology goes beyond purification and equips you with a tool for advanced peptide modification on the solid phase.
Results at a glance
- Benefit from a rapid approach to create purified and modified peptides in a combined approach
- Accelerate development of disulfide peptide drugs
- Access complex modification patterns through PEC-assisted solid-phase modification
Disulfide formation
Disulfide formation remains one of the most attractive modification pathways to improve the properties of peptide drugs. The disulfide bridge provides exceptional chemical and conformational stability. However, manufacturing disulfide peptides is challenging, espe¬cially due to unwanted dimer and multimer formations in solution phase attempts. This case study shows how to create purified disulfide peptides in a combined approach using solid-phase modification with PEC.
The new dimension of purity
Peptide purity is often critically affected by co-eluting side products that are hard to remove by chromatographic methods. Belyntic’s PEC linkers selectively catch your desired product from your crude to reach a new dimension of purity.
Results at a glance
- Discover PEC-grade as a quality label of peptides for preclinical vaccine development
- Synthesize PEC-grade peptides in parallel with unprecedented speed using the 96-well plate format
- Employ PEC-grade peptides in T-cell activation as-says for the development of, e.g., SARS-CoV-2 vac-cines
Peptides vs Virus
The adaption of the PEC technology to a 96 well-plate format gives fast access to purified peptide libraries. PEC removes most peptidic impurities that may cause false-positive signals and improves sub-sequent screening or the reliability of validation assays. These "PEC-grade" peptide libraries show excellent performance in validation studies of SARS-CoV-2 epitopes, likely to elicit a T cell response. This case study highlights the parallel purification's potential during the development of in-silico designed peptide drugs or vaccines.
Results at a glance
- efficiently remove co-eluting truncations in a single step
- get highest purities for challenging peptides with a combination of PEC and RP-HPLC
- save up to 30% of total waste during in orthogonal purification processes
Orthogonality
The purification of the hydrophilic peptide Histone H3 (1-20), synthesized via SPPS, is a formidable challenge due to co-eluting impurities. In this study, we show in collaboration with Bachem how to achieve high purities and lower solvent consumption by using the PEC technology and RP-HPLC in a simple orthogonal purification approach.
Purification of difficult peptides
Solubility and incomplete couplings in lengthy peptides are major issues in peptide manufacturing, limiting exploration and production of novel therapeutics. Belyntic’s catch-and-release methodology enables you to tackle such hard cases and allows you to catch even the most difficult peptides.
Results at a glance
- obtain purified SARS-CoV2 inhibitor peptides with our ready-to-use Research Kit
- apply the PEC routine procedure for PEGylated difficult peptides
- use monodisperse PEG building blocks for further purity optimization
PEGylated Peptides
A recently discovered potential 23-mer peptide therapeutic (SBP1) is a binder to the spike protein of the SARS-CoV-2 virus. Easy access to such peptide sequences with options for further modification is essential to support the global efforts in research labs worldwide to fight against emerging diseases like COVID-19. In this study, we demonstrate the chemical synthesis and purification of N-terminally modified SBP1 with Biotin-(PEG)5 for array applications with our PEC Research Kit. No chromatographic instrumentation is required, also enabling less-equipped lab environments to work with such otherwise inaccessible materials. The modified peptide proved to be functional in the desired assay.
Results at a glance
- purify very long peptides (>50 AAs) in a single step
- isolate peptides from complex mixtures and widen the scope of linear SPPS
- overcome limitations for the manufacturing of hydrophobic or aggregating peptides
Cracking tough nuts
Linear SPPS of long peptides can result in very complex and difficult-to-purify mixtures using liquid chromatography. Also, peptides with hydrophobic or aggregating properties hardly dissolve in media that is compatible with this method. The PEC purification technology can overcome these hurdles, as shown by the purification of very long PTH (1-84) and hydrophobic, strongly aggregating CMV (81-95) peptide.
Higher throughput, higher productivity
Purification is the bottleneck in routine peptide manufacturing in various scales and sequences. Belyntic’s PEC technology finally enables you to processing the purification/modification in parallel and increases your throughput and productivity.
Results at a glance
- significantly improve manufacturing cycles in neoantigen production
- purify neoantigen peptide vaccine with final purities of >90% in a single day
- purify 20 peptides in 3 hours of active working time
20 neoantigens in a single day
The most time-consuming part of the peptide manufacturing process is the purification after synthesis. The PEC technology is a real time-saver at this end. In this case study, we demonstrate the purification of 20 neoantigen peptides with a final average purity of 91% in a single day.
Increased yield
Current purification methods often come with high loss of product. Belyntic’s catch-and-release technology stands out by ensuring a high recovery of product and therefore increased yield for you.
Results at a glance
- purify a wide range of peptides in a routine procedure
- achieve better purities in a single step using PEC in comparison to low-pressure chromatography
- improve the manufacturing economy with higher recoveries
PEC vs. Flash
A collaborated study of Bachem and Belyntic sheds light on the recoveries of the purification of peptides by both Peptide Easy Clean (PEC) technology and reversed-phase flash chromatography. In total, eight peptides were purified with both methods and investigated. The results reveal significant advantages of PEC to improve the yield efficiency of peptide purification.
Routine protocol (one method fits all)
To deal with different peptide properties, the parameters of purification often need to be optimized during laborious method development cycles. Belyntic provides you a simple routine protocol to purify a wide range of peptides.
Results at a glance
- apply optimized TFA-cleavage cocktails to improve crude purities and boost PEC-purities
- we recommend TFA/H2O/PhSMe/EDT/TIS (83:5:5:5:2) or the odorless variant TFA/H2O/DTT/TIS (88:4:6:2)
- suppress the number of tBu adducts in Cys-peptides using Cys(StBu) building block
Scavengers are king
Peptide cleavage and deprotection is a critical step at the end of SPPS. If not performed cautiously, many side products form due to the re-attachment of protecting groups that critically affect crude purity and yield, as well as final purity when using the PEC purification technology. Here we present optimized cleavage cocktails and show how to improve crude and product purity already by choice of amino acid building blocks.
Access to massive parallel purification
Since standard strategies rely on one-by-one purifications, scientists use crude peptides in screenings and discovery processes. Relying on its novel concept, Belyntic’s PEC technology finally holds the key for you to massively parallelize your purification.
Results at a glance
- access massive parallel purification of peptides with PEC in a 96-well filter plate format
- use cost-efficient purified libraries to improve your assay reliability
- shorten the time and cost to prepare purified libraries with PEC
PEC in a 96-well filter plate
The absence of by-products and other impurities in peptide libraries, synthesized via SPPS, is vital for reliable assay results. However, especially in screenings (N > 24), crude peptides are often used because parallel purification of peptides is not feasible. We overcome this bottleneck by customization of our PEC technology to a 96 well-plate format.