We designed the Developer Kit to let purification be integrated flexibly in various workflows and projects. Throughput and scale are adjustable. The kit in elevated bulk scale comes with the essential materials (PEC-Linker, activated filter material) along with a manual.
Developer Kit 10 g: suitable for the purification of approx. 3.3 mmol* peptide
Developer Kit 25 g: suitable for the purification of approx. 8.3 mmol* peptide
Developer Kit 50 g: suitable for the purification of approx. 16.7 mmol* peptide
Developer Kit 100 g: suitable for the purification of approx. 33.3 mmol* peptide
*refers to synthesis scale
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- flexible integration in routine workflow or specialized project scope
- orthogonal to chromatographic purification
- higher productivity through parallel purification
- low solvent consumption
- optional: Please consider our services for implementation or customized protocol development
Note: More consumables are required to run the purification. Please read the FAQ for more information.
|Developer Kit 10||Developer Kit 25||Developer Kit 50||Developer Kit 100|
|PEC-Linker||RC+ (10 g)||RC+ (25 g)||RC+ (50 g)||RC+ (100 g)|
|Activated filter material||Agarose100 ||Agarose100||Agarose100 ||Agarose100 |
|Documentation||Manual (English)||Manual (English)||Manual (English)||Manual (English)|
|Price||€ 2000||€ 4800||€ 8000||€ 15000|
|Documentation||Manual Developer Kit 10||Manual Developer Kit 25||Manual Developer Kit 50||Manual Developer Kit 100 |
|SDS||PEC-Linker RC+ (En)||Mixture of citric acid/sodium carbonate (En)||DL-Dithiothreitol (En)||L-Cysteine (En)|
|Others||Terms and Conditions of Sale||Privacy Agreement|
The PEC-Linker RC+ generation reflects the latest advancements in catch-and-release methodologies (Figure 1). All three construction blocks are optimized and tailored to allow general applicability as well as reliable and highly efficient purification and modification experience for the user.
Figure 1. Molecular structure of the PEC-LInker RC+.
1. Remove the Boc protecting group during the acid treatment for cleavage from the SPPS resin. The activated amino-oxy function serves as the anchor to the activated filter material.
2. A Bromo-substituted para azido-benzyl carbamate acts a the cleavable unit and represents the heart of the PEC-Linker RC+. The construction enables a well-balanced stability behavior, depending on the pH of the medium: Reducing the azide to an amine sensitizes the linker to cleavage. However, the fracture does not occur at neutral pH enabling wash out of by-products formed during reduction. Finally, the treatment of the safety-release system with weak acids liberates the peptide through an acid-catalyzed 1,6-elimination.
3. The para-nitrophenol represents an ideal leaving group with precisely tuned reactivity and storage stability.
The general scheme of PEC purification by catch-and-release consists of six steps shown in Figure 2.
Figure 2. Detailed scheme of the PEC process using the PEC-Linker RC+.
1. Couple the PEC-Linker to the target peptide at the end of the solid-phase peptide synthesis (SPPS). Capping after each amino acid coupling cycle to ensures the selective coupling on the target full-length sequence.
2. Cleave the peptide from the SPPS resin using respective TFA-cocktails.
3. Precipitate and dissolve the peptide.
4. Immobilize through covalent capture ("Catch ") on the activated filter material in an oxime ligation.
5. The covalent capture allows the washing out of unbound substances such as truncated sequences and additionally enables you to modify the bound, unprotected peptide selectively.
6a. The subsequent reduction of the PEC-Linker sensitizes the system for-safety release of the peptide.
6b. Liberate the purified peptide via weak acidic induced 1,6-elimination and elution ("Release").
Aldehyde-modified agarose reflects the state-of-the-art solid-phase equipment for catch-and-release methodologies. We offer an optimized agarose material (Agarose100) with high stability, and a high loading capacity of 100 µmol per mL settled resin in our current kit products.
Figure 3. Microscopy image of the activated agarose filter material.
We point out that the use of Belyntic's kit products with different peptides can lead to very different results due to the peptides' physicochemical diversity. The purification or modification efficiency is also strongly dependent on the synthesis of the peptide itself as well as on the process factors during the use of Belyntic's PEC technology.
Accordingly, it is the responsibility of the user to evaluate the quality of the final product for the intended application. Please conduct suitable analysis and take appropriate measures for further processing and utilization, e.g., by additional purification using chromatographic methods.
Major synthesis requirements are:
- Synthesize peptides by Fmoc-chemistry on the solid phase
- Apply capping after each amino acid coupling step is required
- Suppress re-attachment of protecting groups using decent scavengers
- Avoid aldehyde/ketone contaminations in solvents during precipitation and analysis of crude material
The most important limitations for accessible peptide sequences and design using the PEC-Linker RC+ are:
- N-terminally modified peptides
- disulfides or azides
Please see the FAQ section for more examples of known weaknesses and sources of contamination according to the current knowledge of Belyntic.
“For the first time, the Belyntic PEC Kit enabled us to provide peptides that we couldn't purify before.”
Dr. Adrian Glas
Peptide Specialty Laboratories GmbH (PSL)
“[…] our collaboration has highlighted the potential of PEC in either single-step purification or as a pre-LC clean-up step for very complex crude peptide mixtures, making it a valuable addition to our toolbox. The flexible nature of the PEC purification method also provides an additional platform for more unusual peptide chemistry […]”
Dr. Gavin Noble
“With Belyntic's PEC technology, a set of 48 short peptides were rapidly provided, that we were able to use immediately for the development and validation of LC-MS/MS quantification methods. Their team was extremely helpful and responsive. They helped us move a novel project forward very quickly.”
Dr. Alex Kentsis
Memorial Sloan Kettering Cancer Center
“[…] The time saved is precious, enabling technicians to work on other tasks. Also, Belyntic has offered fantastic customer support, with fast communication and great suggestions on how to solve issues to achieve the highest yield and purity.”
Dr. Jonas E. Jensen
“The PEC linker is a significant new invention in peptide synthesis technology, and every peptide chemist can benefit from it. PEC enables all peptide-based biopharmaceutical businesses to considerably increase efficiency in terms of speed and cost of goods, and we expect it to become a standard method of art in peptide production technologies.”
Dr. Thomas Bruckdorfer (CSO & VP Business Development)
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 sho...Go to case study
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 sim...Go to case study
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 dise...Go to case study
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.Go to case study
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 prese...Go to case study
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 customi...Go to case study