With over 20 years of experience in peptide synthesis, we have advanced peptide synthesis instruments, professional team, and extensive expertise in peptide synthesis and purification processes. Our advanced peptide synthesis platform and quality control system work together to provide clients with high-quality products.
In peptide synthesis, when a peptide is reported as being 95% pure, it means that 95% of the material in the sample consists of the desired peptide sequence. The remaining 5% is composed of impurities or related substances. These impurities can arise from various sources during the synthesis, purification, and handling processes. Common impurities in peptide synthesis include:
Incomplete Coupling Products: These are peptides where not all amino acids have successfully coupled together during the synthesis, leading to shorter-than-desired sequences.
Deletion Sequences: These are peptides where one or more amino acids are missing in the sequence due to incomplete reactions during the synthesis.
Side-Chain Protecting Group Adducts: The protecting groups used during the synthesis may not be completely removed, leading to adducts with the peptide.
By-Products of Coupling Reactions: Some reactions may produce unwanted by-products that can contribute to impurities in the final product.
Residual Reagents: Traces of reagents used in the synthesis or purification steps may remain in the final product.
Analytical techniques such as high-performance liquid chromatography (HPLC) and mass spectrometry (MS) are commonly employed to assess the purity of synthesized peptides and identify the nature of impurities present. These techniques allow researchers to characterize the composition of the peptide sample and make adjustments to the synthesis or purification process if necessary.
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Peptide purification is a crucial step in the production of high-quality peptides for various applications, such as research, pharmaceuticals, and diagnostics. The most common method for peptide purification is high-performance liquid chromatography (HPLC), specifically reverse-phase HPLC (RP-HPLC). Here's an overview of the peptide purification process:
Solid-Phase Peptide Synthesis (SPPS):
Cleavage and Deprotection:
Crude Peptide:
Purification by RP-HPLC:
Analysis and Characterization:
Optional Additional Steps:
Lyophilization:
It's important to note that the purification strategy may vary based on the characteristics of the peptide, the scale of synthesis, and the intended application. Additionally, automated peptide synthesizers and purification systems are commonly used to streamline and optimize the process.
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$16.5Mn in financing, including a $7.5Mn Series A+ round led by Engine Ventures with participation from Forcefield Venture Fund. It previously raised a $3.55Mn Series A round led by Engine Ventures and a $5.45Mn Seed round led by Biological Engineering Ventures.$92.5Mn for the facility and associated assets, subject to customary adjustments in accordance with the agreement.November 2023
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The Anhui Provincial Peptide Drug Engineering Laboratory (University of Science and Technology of China), in collaboration with a team from Nanjing University, has reported the latest advances in the field of asymmetric radical acylation achieved through photoenzyme catalysis in the journal “Nature.”
In recent times, the team led by Professor Tian Changlin from the Anhui Provincial Peptide Drug Engineering Laboratory (Biomedical Department, University of Science and Technology of China, and the High Magnetic Field Science Center, Chinese Academy of Sciences) collaborated with Professor Huang Xiaoqiang’s team and Professor Liang Yong’s team from Nanjing University to make significant strides in the field of photoenzyme catalysis.
In response to the developed dual catalytic system involving thiamine diphosphate (ThDP)-dependent enzymes and photocatalysis using phosphorus-amino acid (ThDP) as the catalyst, various reaction intermediates, such as free radicals in many reaction processes, changes in the oxidation state of metal catalysts involved in the catalytic reaction, and electron transfer processes during oxidation-reduction, were identified and analyzed using electron paramagnetic resonance (EPR) methods. Professor Tian Changlin’s team at the School of Life Sciences, University of Science and Technology of China, has long been engaged in research at the High Magnetic Field Science Center of the Chinese Academy of Sciences, focusing on the identification of free radicals and analysis of electron transfer in research related to high-field EPR equipment setup, low-temperature EPR method development, and the mechanisms of chemical catalysis and enzyme catalysis, achieving a series of research results (Nat Catalysis 2023; Angew Chem Int Ed 2023, PNAS, 2023, 2022; ACS Catalysis 2023, 2021; Chem Commun, 2022, 2021; Science 2018, etc.). Recently, Professor Tian Changlin’s team collaborated with Professor Huang Xiaoqiang’s team and Professor Liang Yong’s team at Nanjing University to make significant progress in the field of photoenzyme catalysis. Using EPR methods, they identified the free radical intermediates in the newly developed catalytic system and the electron transfer mechanism in the catalytic reaction. The research results, titled “A light-driven enzymatic enantioselective radical acylation,” were published in Nature (DOI: 10.1038/s41586-023-06822-x).
Biomanufacturing is one of the most promising green technologies for transforming industrial sustainability and is a core aspect of enzyme catalysis in synthetic biology. The combination of enzyme catalysis and photocatalysis, known as photoenzyme catalysis, integrates the diverse reactivity of photochemistry with the high selectivity of enzymes, making it the forefront strategy for developing new enzyme functions. The collaborative research team, using a combination of biomimetic and chemical simulation approaches (Figure 1), harnessed visible light excitation and directed evolution to extend enzyme catalytic functions to radical-radical cross-coupling. Additionally, by using directed evolution to modify ThDP-dependent enzymes, they reshaped ThDP-dependent benzaldehyde lyase into a radical acyl transferase (RAT), achieving a non-natural high enantioselective radical-radical coupling reaction.
The collaborative team explored the catalytic system of organic dye Rose Bengal and ThDP-dependent enzyme using 4-methoxybenzaldehyde 1a and free radical precursor oxidation-reduction active ester 2a as template substrates. Subsequently, a small and refined mutant library was constructed through molecular dynamics simulations and semi-rational design. The optimal mutant enzyme with high substrate tolerance and substrate selectivity (enantioselectivity up to 97% ee) was obtained, highlighting the finely tuned role of the enzyme’s adjustable active pocket in the stereochemical control of free radical stereochemistry (Figure 2).
For the photoenzyme dual catalytic system, Professor Tian Changlin’s team applied low-temperature (80K) electron paramagnetic resonance (EPR) experiments, capturing the ThDP-derived ketyl free radical (Int. B). Through EPR spin trapping experiments, they detected characteristic six-line splitting spectra in the standard reaction system, confirming it as an intermediate benzylic radical (Int. C) and the free radical product after addition with the capture agent. This provided direct evidence for unraveling the key to the new enzyme reactivity and the source of high stereochemical selectivity.
The collaborative development of a dual catalytic system combining ThDP-dependent enzyme catalysis and organic photosensitizer Eosin Y catalysis, led by Nanjing University and involving the team from the University of Science and Technology of China, not only transformed natural benzaldehyde lyase into a light-driven radical acyl transferase but also achieved excellent stereochemical control of a challenging prochiral free radical. Nanjing University is the first and last corresponding author unit, and the University of Science and Technology of China and the Anhui Provincial Peptide Drug Engineering Laboratory are co-corresponding author units. The aforementioned research work received funding from the National Natural Science Foundation of China’s Outstanding Youth Fund, major instrument development projects, and the Ministry of Science and Technology’s key research and development program.
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On December 8th, the U.S. Food and Drug Administration (FDA) approved two groundbreaking therapeutic drugs, Casgevy (developed jointly by Vertex Pharmaceuticals and CRISPR Therapeutics) and Lyfgenia (developed by Blue Bird Bio). These drugs represent the first wave of cell-based gene therapies approved by the FDA for the treatment of sickle cell disease (SCD) in patients aged 12 and above. Notably, Casgevy is the first FDA-approved therapeutic drug utilizing CRISPR gene editing technology, marking a significant milestone in the field of gene therapy.
It’s worth mentioning that Lyfgenia’s parent company, Blue Bird Bio, received approval for another drug called Zynteglo in August 2022, intended for the treatment of transfusion-dependent β-thalassemia (TDT).
The structural composition of human hemoglobin consists of a heterotetramer, composed of two pairs of globin proteins. SCD and TDT result from variations in the β-globin protein. SCD involves a mutation where Glu6 in β-globin is replaced by Val6, causing hemoglobin to form long chains through hydrophobic interactions, leading to sickle-shaped cells and reduced oxygen-carrying capacity. TDT results from deficient synthesis of the β-globin chain, causing an excess of unbound α-chains, leading to precipitation and hemolysis within red blood cells.
Both treatments involve extracting bone marrow hematopoietic stem cells from the patient, editing them in the laboratory using either CRISPR or viral vectors, and then reintroducing the edited stem cells into the patient.
While Casgevy and Lyfgenia represent significant milestones in the treatment of SCD, certain factors limit them from being the ultimate solutions. Safety concerns, potential carcinogenic risks associated with viral vectors (as in the case of Lyfgenia), and the long-term safety of gene editing are issues that need further investigation. Both therapies also involve myeloablation (bone marrow conditioning), leading to immune and reproductive challenges.
Another critical factor is the pricing. With Casgevy priced at $2.2 million and Lyfgenia at $3.1 million, the high costs raise questions about accessibility and fairness, especially for SCD patients in sub-Saharan Africa. Traditional treatments, while not curative, are significantly more affordable.
In conclusion, Casgevy and Lyfgenia are indeed milestones, but they are not the endpoint for SCD. The emergence of gene editing therapies marks the beginning of a new era, providing hope for revolutionary treatments. However, challenges such as safety concerns, accessibility, and pricing highlight the need for continuous advancements and ethical considerations in the evolving landscape of gene therapies. As we witness these groundbreaking treatments, Winston Churchill’s words come to mind: “Now this is not the end. It is not even the beginning of the end. But it is, perhaps, the end of the beginning.” The advent of these therapies signifies a new chapter, and while celebrating their potential, we must also address the challenges and questions they bring.
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