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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Highlights:

- 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


The PEC-Linker RC+


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.



PEC-Linker RC+ in action


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").



The activated filter material


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.

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