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Chem-Enzymatic Peptide Synthesis (CEPS) Technology

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Chem-enzymatic peptide synthesis (CEPS) technology provides a flexible and more specific method for peptide synthesis. It is able to handle a variety of long and complex peptides and is the simplest method for synthesizing long peptides in active conformations without the need for recombinant production. CD Formulation uses advanced CEPS technology to provide high-quality, custom peptide synthesis services tailored to meet the specific needs of researchers and pharmaceutical companies. With CEPS technology, we can produce peptides that maintain their biological activity and desired conformation to meet the needs of different customers.

What is Chem-Enzymatic Peptide Synthesis (CEPS) Technology?

Large-scale chemical production of long peptides remains a huge challenge. Using chemical methods such as solid-phase peptide synthesis (SPPS), synthesis yields decrease significantly with increasing peptide chain length. Crude purity and overall yield can be significantly improved using fragment condensation strategies. However, chemical fragment condensation leads to epimerization (except for Gly and Pro residues), native chemical ligation is often not feasible (Cys is not present), and scale-up is difficult due to thioester instability. Furthermore, recombinant expression of peptides as an alternative method cannot guarantee the solubility of the final product. As a cutting-edge technology, CEPS has a unique multi-functional engineering enzyme that can achieve the conversion of non-side chain-protected polypeptides with absolutely no sidechaining. It is the easiest way to deal with long and complex peptides.

Fig. 1 Chemo-Enzymatic Peptide Synthesis (CEPS) method. Fig. 1 Overview of the Chemo-Enzymatic Peptide Synthesis (CEPS) method. (Liu Y, et al., 2022)

Our Services Related to Chem-Enzymatic Peptide Synthesis (CEPS) Technology

CEPS is a hydrolase-catalyzed stereoselective formation of peptide bonds. Compared to traditional chemical synthesis with harsh reaction conditions, it is a clean and mild procedure without the need for complex and laborious protection-deprotection procedures. The technology is capable of processing long and complex peptides and is the simplest way to synthesize long peptides in active conformations without the need for recombinant production.

Our team has extensive experience in processing long-sequence peptides and cyclic peptides using advanced CEPS technology and can synthesize peptides up to 160-200 amino acids in length.

Commonly Used Enzymes in Chem-Enzymatic Peptide Synthesis (CEPS)

Enzyme Types	Name	Description
Cysteine Proteases	Papain	Papain has broad substrate specificity and is stable and active from pH 4 to pH 10 and at high temperatures (up to 80 °C). Additionally, papain uses l-glutamate and various amino acid esters as substrates to achieve protease-catalyzed amino acid copolymerization.
Cysteine Proteases	Bromelain	Bromelain has broad specificity for cleaving proteins, especially for polar amino acids at the P1 position.
Serine Proteases	α-chymotrypsin	The main cleavage site of α-chymotrypsin is the peptide bond, and the carboxyl side chain is a hydrophobic amino acid such as tyrosine, tryptophan, and phenylalanine.
	Protease K	Proteinase K, an enzyme of remarkable versatility, exhibits an astonishingly expansive substrate specificity. It slices through peptide bonds adjacent to the carboxyl group of both aliphatic and aromatic amino acids with mercurial finesse, and it can orchestrate the hydrolysis of esters. This proteolytic powerhouse thrives in a remarkably diverse pH milieu, spanning from 7.5 to an alkaline 12.0, and maintaining activity at elevated temperatures reaching a sweltering 60°C.
	Trypsin	Trypsin is specific for the C-terminal peptide bonds of lysine and arginine residues, with the exception of -Arg-Pro- and -Lys-Pro- bonds.
	Subtilisin	Subtilisin has good stability in the pH 7-10 range, and the subtilisin catalytic strategy is established in pure organic solvents.
Esterases	Lipase	Lipases can catalyze the hydrolysis of triglycerides as well as esterification, amidation, and transesterification reactions.

Advantages of Chem-Enzymatic Peptide Synthesis (CEPS) Technology

CEPS is a clean and gentle procedure that does not require complex and laborious protection-deprotection procedures.
CEPS can generate longer peptides than SPPS, with a length of 160-200 amino acids.
CEPS yields more than two times higher than SPPS, making it an excellent alternative to traditional chemical peptide synthesis.
Pre-purified fragments are used for ligation to obtain higher purity products than SPPS.
CEPS's chemical selectivity allows the conversion of non-side chain-protected peptides and the absence of side chaining.
CEPS can produce new peptide variants quickly and with a high success rate.
CEPS can significantly reduce the costs associated with peptide synthesis.

Custom Chem-Enzymatic Peptide Synthesis (CEPS) Services

Therapeutic Peptide Synthesis and Production

Many peptide drugs are developed by chemically modifying natural molecules or by artificial synthesis. CD Formulation has extensive experience in custom therapeutic peptide synthesis and production. Each synthesis step is subject to our strict quality control. We can produce various synthetic peptides according to customer requirements, from simple to long and challenging sequences. Leverage our best-in-class CEPS technology to support the synthesis of your target peptides.

Deliverables

CD Formulation's peptides are delivered as lyophilized powders and packaged in separate sample tubes according to customer requirements. To ensure the quality of the peptides, we also provide the following documents for your reference:

HPLC/MS Test: All peptides we produce are tested by MS and HPLC to ensure the accuracy of the peptide products.
COA Document: We provide a COA document for each peptide, including the shape and appearance of the peptide sample, delivery specifications, molecular weight, and purity.
Solubility Test: This is optional. We provide two solubility testing services according to customer requirements: qualitative dissolution test and quantitative dissolution test.

Publication

Published Data

Technology: Chem-Enzymatic Peptide Synthesis (CEPS)

Journal: Int J Biol Macromol.

IF: 7.7

Published: 2023

Results:

The authors designed and developed a CEPS process for the synthesis of aviptadil, a 28-mer therapeutic peptide. Peptiligase was used as the catalytic enzyme. The results showed that the peptide ligase variant was able to complete the ligation reaction within 15 minutes. The total ligation time was shorter than that of peptides of similar length and similar hindrance to aviptadil.

CD Formulation is designed to provide powerful tools for handling the synthesis of peptides of varying lengths and complexities. Please feel free to contact us if you are interested in our services. Learn how our CEPS technology can support your custom peptide synthesis projects.

References

Liu Y, Güler R, Liao Y, et al. Biologic Evaluation of a Heterodimeric HER2-Albumin Targeted Affibody Molecule Produced by Chemo-Enzymatic Peptide Synthesis. Pharmaceutics. 2022;14(11):2519.
Mahmoudzadeh K, Habibi Z, Yousefi M, et al. Peptiligase, an enzyme for efficient chemo-enzymatic synthesis of aviptadil. Int J Biol Macromol. 2023;253(Pt 7):127089.

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