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.
![PEC-Linker RC+](https://www.gyrosproteintechnologies.com/hs-fs/hubfs/GPT/pdfs/1-PTI/PurePep%20EasyClean%20Purifications%20Kits/images/PEC-Linker%20RC+.png?width=400&height=123&name=PEC-Linker%20RC+.png)
Figure 1. Molecular structure of the PEC-LInker RC+.
- 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.
- 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.
- 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.
![Detailed scheme of the PEC process using the PEC-Linker RC+.](https://www.gyrosproteintechnologies.com/hs-fs/hubfs/GPT/pdfs/1-PTI/PurePep%20EasyClean%20Purifications%20Kits/images/Detailed%20scheme%20of%20the%20PEC%20process%20using%20the%20PEC-Linker%20RC+..png?width=1000&height=149&name=Detailed%20scheme%20of%20the%20PEC%20process%20using%20the%20PEC-Linker%20RC+..png)
Figure 2. Detailed scheme of the PEC process using the PEC-Linker RC+.
- 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.
- Cleave the peptide from the SPPS resin using respective TFA-cocktails.
- Immobilize through covalent capture ("Catch ") on the activated filter material in an oxime ligation.
- 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.
- Precipitate and dissolve the peptide
- a) The subsequent reduction of the PEC-Linker sensitizes the system for-safety release of the peptide.
b) 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.
![Microscopy image of the activated agarose filter material](https://www.gyrosproteintechnologies.com/hs-fs/hubfs/GPT/pdfs/1-PTI/PurePep%20EasyClean%20Purifications%20Kits/images/Microscopy%20image%20of%20the%20activated%20agarose%20filter%20material%20(1).png?width=400&height=300&name=Microscopy%20image%20of%20the%20activated%20agarose%20filter%20material%20(1).png)
Figure 3. Microscopy image of the activated agarose filter material.