Our High-Throughput Kits open doors for the massive parallelization of peptide manufacturing. Create purified peptide libraries using catch-and-release purification methodologies in 48- or 96- well filter plates. The high loading capacity of the activated filter material allows you to address the preparative scale with unprecedented speed.

High-Throughput Kit 96 x 10 µmol:        Get access to a purified peptide library in  preparative scale within a day

High-Throughput Kit 48 x 20 µmol:        Seamlessly adjust your scale and throughput for pre-clinical studies

Highlights:

- parallel purification in 48- or 96-well filter plates

- high-loading capacity purification media allows scales up to 20 μmol

- increase the reliability of your screening or validation assay

- acrylic vacuum manifold available in the webshop

*referring to synthesis scale

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Note: More consumables are required to run the purification. Please read the FAQ for more information.

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 

- 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.