of solid phase peptide synthesis
The peptide synthesis team at Enzene has extensive experience in the development of peptide products from milligram to gram scale. We have various platforms, which include liquid phase, solid phase and automated peptide synthesis. Our activities include designing novel synthesis and purification technologies for peptides. Developing long chain peptides (upto 40 aa), modified peptides, cyclic peptides, disulphide bridged peptides and complex peptide mixtures. We also have various modification platforms PEGylation, Biotinylation, FITC labeling, phosphorylation, methylation, acetylation, amidation & conjugation. We have successfully transferred peptide synthesis process of up to kilogram scale for GMP manufacturing of therapeutic peptides.
Synthesis of peptides by solid phase method.
The Group-Assisted Purification (GAP) strategy has been successfully utilized for the solution phase synthesis of N-protected amino acids and peptides by avoiding the use of traditional purification protocols such as chromatography and recrystallization. In the present GAP synthesis, pure products have been obtained simply by washing the crude mixtures with inexpensive petroleum solvents or co-solvents to give good to high yields. In addition, the GAP auxiliary can be conveniently recovered for re-use.
Recently, our labs have established the GAP (Group-Assisted Purification) chemistry concept through the design and synthesis of new reagents that were attached with unique achiral and chiral auxiliaries.– By using GAP technology, organic synthesis can be performed efficiently and stereochemically without the use of traditional purifications of chromatography, recrystallization, etc. In fact, the stereoisomeric products can be obtained simply by washing the crude mixtures with inexpensive petroleum solvents or co-solvents. Therefore, it can substantially reduce the use of starting materials, silica gels, energy, manpower, etc. In addition, achiral and chiral auxiliaries can be conveniently recovered for re-use; and they can often be quantitatively recovered via a one-time extraction with n-butanol. It is believed that the GAP chemistry concept will have a huge impact on chemical synthesis and medicinal fields. In this report, we would like to disclose that the GAP chemistry strategy can advance peptide synthesis, which has advantages of both solid-phase-peptide synthesis (SPPS) and liquid-phase synthesis of peptides by avoiding their shortcomings in regard to the factors mentioned above.–
Protocol for Peptide Synthesis and Self-Assembly
Although there have recently been significant advances in solid phase peptide synthesis, most notably the virtually universal transition from t-Boc to FMOC based chemistries, peptide synthesis still remains somewhat of an art, definitely not a routine process. In the past, particular amino acids or combinations of amino acids posed significant obstacles for synthesys, as did longer peptides. The new chemistries have shifted the synthetic problems to primarily sequence-based ones. While this has allowed the more routine synthesis of short peptides, and increased the potential for successful synthesis of longer peptides, prediction of problem regions is still often elusive. This makes stringent quality assurance measurements for every peptide synthesized at the PRC an essential part of the synthetic process. Each peptide delivered to an investigator is fully validated by LC-MS (liquid chromatography-mass spectrometry).
An introductory guide to solid phase peptide synthesis
Synthetic peptides have a wide variety of uses ranging from structure-function analysis of sites within the protein, binding assays, receptor agonists/antagonists, to immunogens for the production of antisera. Regardless of the specific application, all that is required from the investigator to initiate peptide synthesis is the sequence of the desired peptide and an account number. We will attempt to synthesize virtually any sequence provided by the investigators. The Peptide Synthesis Service provides a complete range of services including cleavage, HPLC purification, quality assurance analysis and a variety of amino acid derivatization/labeling techniques.
peptide cleavage from resins, peptide purification and storage ..
Since the purity of the peptide obtained from the synthesis is sequence dependent, purification to >95% will be performed upon request. Major impurities can range from 10% to 50% by weight. They consist of small water-soluble molecules, salts and protecting groups from the cleavage reaction, deletion peptides created due to incomplete coupling during synthesis, and modified peptides created during the cleavage. These species can be removed using reversed phase HPLC. We recommend that, under most circumstances, all peptides be purified prior to employing them in research studies.