Peptide Synthesis Methods: Traditional Vs. Modern Approaches
There are several essential components to the biological and chemical processes of life. On a cellular level, there are certain building blocks or compounds that, as their name implies, construct the framework of life.
When these substances are combined with other substances, they serve an essential functional purpose.
Thanks to molecular research and biochemistry advancements, it is now possible to produce or combine chemicals in a lab and use them for diverse purposes in medicine, industry, pharmaceuticals, etc.
The development of recombinant proteins, bespoke proteins, and other cutting-edge techniques has become increasingly common in science and research. Peptide Synthesis is one such process
Contents
What are Peptides?
Peptides are extensive chains of amino acids found in the human body. Although amino acids are used to produce proteins, peptides contain fewer proteins than proteins. Peptides, identical to those in your body, can also be made in laboratories.
Recombinant protein creation, synthesis, and other cutting-edge methods are now widely used in science and research. Recombinant protein expression is a service that is provided in addition to protein expression and purification services.
Many commercial gene and peptide synthesis firms, along with monoclonal antibody sequencing, are highly used for their application in the pharmaceutical industry.
In addition to researching Elisa kits, e-coli protein aggression, and antibody sensitivity, these commercial agents provide peptide synthesis services.
Peptide therapy, when integrated into their service offerings, allows these commercial agents to also address therapeutic applications, enhancing treatment options for a variety of health conditions through tailored peptide solutions.
You must grasp the biological properties of amino acids, especially those best suited for peptide synthesis before you can comprehend the peptide synthesis process. For the best outcomes, researchers specializing in peptide synthesis or protein production and purification employ these amino acids.
One such compound is a proteinogenic amino acid. They are renowned for serving as proteins’ building blocks. They are biosynthetically added throughout the expression or synthesis process. Proteinogenic means “protein creating,” as the name suggests.
How are Peptides Synthesised?
As previously mentioned, amino acid chains are assembled to form peptides. During the production process, the carboxylic acid of one amino acid joins the amine group of another acid. The R group, a distinct functional group that gives each amino acid uniqueness, is present in all amino acids.
This functional R group frequently controls a peptide’s general shape, structure, and property. To completely understand the complexity of peptide creation, we must first understand the role that amino acids play, particularly when they are used to synthesize peptides.
The amine group of amino acids must be appropriately protected during synthesis to avoid unexpected reactions. This is done by using a chemical called FMOC.
The two methods of peptide production are:
- Liquid phase peptide synthesis:
- Solid phase peptide synthesis (SPPS)
We’ll discuss these methods elaborately below:
Solid Phase Peptide Synthesis
The Solid Phase Peptide Synthesis (SPPS), created in 1963 by Nobel Prize–winning chemist Robert Bruce Merrifield, has grown to be the most used peptide synthesis approach.
The attachment of each peptide chain to polystyrene beads is the essential component of SPPS.
Through this procedure, it is now possible to investigate a peptide inside a vessel. The vessel uses a filter to keep the peptide-connected resin in place, allowing solutions to drain afterward.
An amino acid that is already bonded to a resin catalyzes the process. Additionally, the subsequent phases typically look something like this:
- Removal of FMOC from the amine side group of amino acids
- Addition of the following amino acids in the strain
- Activation of coupling reagents
- Repetition of the first three steps until completion of the sequence
- Cleaving the peptide from the resin
It’s important to remember that each FMOC group protects the amine side of an amino acid. As mentioned, two amino acids are joined together by coupling the NH2 side of one with the COOH side of another. SPPS
- FMOC
One of the best processes for synthesizing peptides is the FMOC SPPS. High-quality FMOC building blocks are widely available and reasonably priced. Depending on the kind of synthesis, some FMOC blocks also have changes supporting peptide generation.
Conversely, as peptides’ commercial potential has grown, so too has the demand for FMOC uses, which has accelerated those applications’ technological development.
- Resin Cleavage
Removing all side chain protective components utilized during the peptide synthesis process, particularly the detachment of the peptide from the supporting resin, is a vital step once the synthesis process has been finished using the FMOC SPPS method.
A damaged, freshly synthesized peptide and the bonding of amino acids with other amine group molecules are likely if this task is left unfinished. Deprotection is another name for this procedure. The peptidyl resin is often cared for using TFA when using the FMOC SPPS system.
- Peptide Purification
Removing undesired elements or residual effects from the amino acid coupling can be one of the most time-consuming steps in the peptide synthesis process. Identifying unwanted substances is challenging since they resemble the intended products closely. Following the treatment, the following contaminants are frequently discovered:
- Deletion peptides
- Truncated peptides
- Scavengers
- By-products of protection elements
The identification technique must be because the impure ingredients and the result are identical. Searching for contaminated materials using reversed-phase HPLC with UV peak detention can be done.
However, ensure that the contaminants are eliminated throughout this process and that the synthesized peptide’s natural properties are not altered.
Liquid Phase Peptide Synthesis
The liquid phase was the predominant peptide synthesis technique before SPPS was discovered. It was previously referred to as the standard organic chemistry solution.
After discovering that peptide could be made using the LPPS method, American biochemist Vincent du Vigneaud also received the Nobel Prize in Chemistry.
However, the SPPS approach has a far higher potential for peptide synthesis. The conventional nature of peptide synthesis is still used for large-scale manufacturing of peptides, particularly for industrial uses.
Conclusion
Understanding the complexities of molecular substances and their conventional responsibilities in your body, such as development, healing, and energy supply, requires a critical biological method called custom peptide synthesis.
With purification, the synthesis process has improved dramatically in both accessibility and efficacy, especially regarding peptide purity.
These services are being increasingly utilized by peptide synthesis firms for additional study into the field so that new improvements may be produced.