PEPTIDES & PROTEINS
Fermentation process & sustainability
Microbial fermentation for the production of peptides and nucleotides is considered a sustainable process for several reasons . First , it is inherently safe and energy-efficient . Water serves as the primary solvent and the temperatures typically remain below 50 ° C . The reagents used in the process are mostly safe and fermentation mainly involves providing sugars to microbes .
All this aligns with several Green Chemistry principles related to safety and low energy consumption . Additionally , the process utilises renewable resources , as both the microorganisms and the reagents are renewable , with water being the main solvent ( Figure 2 ). This is in compliance with the principles that emphasise renewable inputs .
Peptide synthesis in microbes relies on natural enzymes , which act as catalysts , eliminating the need for stoichiometric reagents or protection steps . This contributes to maximising atom efficiency , as no wasteful intermediates are produced . The process thus adheres to principles related to catalysis and atom economy .
Furthermore , the waste generated from microbial fermentation is mostly water and non-toxic materials , and the final by-products , such as degraded peptides , can be safely released into the environment . This aligns with the principle of waste prevention , as the waste is minimal and biodegradable .
The materials used in microbial fermentation are biodegradable , which supports the principle of using materials that degrade after use . Overall , microbial fermentation for peptide production is largely in line with the core principles of Green Chemistry , making it a ‘ green ’ technology . However , there is still room for improvements to further enhance its sustainability .
Regulatory challenges
One of the main challenges of recombinant technology lies in regulatory hurdles . The FDA and EMA
Water
26 %
1 %
Raw materials
73 %
Raw aterials Figure 2 - Composition of process mass intensity from microbial fermentation industrial processes
Consumables
guidelines are not fully applicable to recombinant biotechnological products , mainly due to concerns about the potential immunogenicity of impurities from host cell proteins . These residual elements , such as host proteins and DNA , must be carefully monitored , quantified and controlled to ensure product safety .
To analyse host cell impurities in recombinant peptide production , several advanced techniques are employed . One such technique is ELISA ( enzyme-linked immunosorbent assay ), which is highly sensitive and allows for the detection and quantification of HCPs by utilising antigen-antibody interactions .
Another is 2D-DIGE ( two-dimensional difference gel electrophoresis ), which separates proteins based on their size and charge . This separation helps identify HCPs by comparing samples under different conditions . Mass spectrometry is also a key tool in this process , as it identifies and quantifies proteins by analysing their massto-charge ratio , providing detailed information about the composition of impurities .
In silico epitope prediction employs computational methods to forecast potentially immunogenic regions in proteins , aiding in the identification of harmful epitopes related to HCPs . Finally , in vitro assays simulate biological conditions to assess the functional impact of impurities and determine their potential to trigger immune responses . By using these techniques together , a thorough monitoring and control of HCPs is achieved , ensuring the purity and safety of recombinant peptide products .
Despite these advances , challenges persist due to the absence of unified regulatory guidelines for peptide drugs . The term peptide does not have a standardised regulatory definition , and the regulatory pathway for each product is primarily shaped by historical precedents .
These analytical tools play a crucial role in ensuring the purity and safety of the final product , which is essential for supporting regulatory submissions . While regulatory authorities have made notable progress , additional work is required to establish comprehensive guidance on impurity control strategies and safety standards for recombinant peptide products .
Regulatory pathways also play a significant role in the choice of production method . Some techniques , particularly those involving recombinant DNA or biological systems , may require more stringent regulatory oversight , especially regarding manufacturing practices and product consistency .
Conclusion
Selecting the right manufacturing technique for peptides involves balancing its characteristics , scalability needs , cost considerations and regulatory requirements . Each approach has its own set of challenges , but careful evaluation of these factors will help identify the most efficient and viable solution . Recombinant peptide production stands out as a promising solution to help address this challenge . ●
J j
Maurizio Sartorato
VICE PRESIDENT BIOTECH
OLON msartorato @ olonspa . it www . olonspa . it
38 SPECIALITY CHEMICALS MAGAZINE ESTABLISHED 1981