Proteins are the building blocks of life and play a decisive role in biological processes. The following examples illustrate their importance and diversity:
This diversity requires coordinated biosynthesis. The basic building blocks of proteins are amino acids, organic compounds that are strung together like pearls (specific to each protein). The amino acid sequence (primary structure) of a protein is genetically determined in the DNA sequence of an organism. However, proteins only achieve structural stability through interactions between non-adjacent amino acids and thus through folding into higher-level, three-dimensional structures (tertiary structure). In addition to intramolecular interactions (monomer – interaction of amino acids of one protein molecule), intermolecular interactions (dimer, trimer, multimer – interaction of amino acids of two, three or more protein molecules) lead to the formation of complex quaternary structures. These three-dimensional structures are specific to each protein of an organism and are essential for its functionality. Even small errors can lead to a complete loss of function.
The complexity of proteins makes them interesting objects for research and development, both in basic research and in the development and production of biotherapeutics. Particle size, zeta potential/charge, molecular mass, microrheological properties and stability are important and essential protein parameters.
3P Instruments offers instrumental solutions for various questions in protein analysis, such as
Structural and functional analyses of proteins are only possible by isolating and enriching them from biological systems. The optimisation of the purification process is an important prerequisite for the stability of an isolated protein, whose in vivo (= synthesised in the organism) integrity and complexity must also be maintained in vitro (= isolated protein). Cell disruption, temperature, pH, salt content, ageing through storage and other stress factors lead to changes in the physical and chemical properties and thus to denaturation of the protein through destruction of the tertiary and/or quaternary structure or through aggregate formation. The identification of such denaturation processes is the key to stabilised, isolated proteins for structural and functional analyses. The integrity of an isolated protein is only guaranteed under optimal conditions.
3P Instruments offers various options for protein characterisation.
| Area | Target | Task | Parameter | Method | Device |
| Protein isolation and characterisation | Purified protein | Characterisation | Particle size | DLS | BeNano |
| Molecular mass | SLS | ||||
| Zeta potential | ELS | ||||
| Microrheology | µ-DLS | ||||
| (Thermal) stability/lability | Particle size | DLS | BeNano (+ BAT-1) | ||
| Particle size vs. temperature | DLS | ||||
| Zeta potential vs. temperature | ELS | ||||
| Zeta potential vs. pH | ELS | ||||
| Microrheology | µ-DLS | ||||
| Dispersion stability | Scan | ||||
| Structural changes | DSC |
With the help of protein engineering, proteins can be genetically synthesised in homologous or heterologous systems (recombinant proteins). The focus of targeted expression is usually on increasing yield and simplifying purification. The identity between native and recombinant protein is important here. Structural differences often lead to a loss of stability and/or function. Structural changes in proteins can also be caused by mutations, which can lead to a loss of function but also to increased functionality (e.g. enzyme activity). In addition to naturally occurring mutations (faulty recombinant biosynthesis in vivo), this applies in particular to mutations specifically introduced by genetic engineering. Different analytical methods help in the comparative protein characterisation for the establishment of recombinant expression systems.
3P Instruments offers numerous options for analysing recombinant proteins with regard to integrity and stability, in order to uncover possible structural and, consequently, functional differences.
| Area | Target | Task | Parameter | Method | Device |
| Recombinant proteins (protein expression homologous/ heterologous) | Recombinant protein | Characterisation | Particle size | DLS | BeNano |
| Molecular mass | SLS | ||||
| Zeta potential | ELS | ||||
| Microrheology | µ-DLS | ||||
| (Thermal) stability/lability | Particle size | DLS | BeNano (+ BAT-1) | ||
| Particle size vs. temperature | DLS | ||||
| Zeta potential vs. temperature | ELS | ||||
| Zeta potential vs. pH | ELS | ||||
| Microrheology | µ-DLS | ||||
| Dispersion stability | Scan | ||||
| Structural changes | DSC |
Isolated native or recombinant proteins are frequently used as biotherapeutics. On their way from the laboratory to the patient, these therapeutics pose particular challenges due to their instability. At every stage of the development process, it must be ensured that the products fulfil the requirements (safety, efficacy, user-friendliness). The establishment of stable production, storage and transport conditions is an essential prerequisite for preventing protein agglomeration/aggregation and, as a result, flocculation (risk of syringe clogging) and ensuring functionality (e.g. ligand binding, enzyme activity, etc.). Different analytical methods help with comparative protein characterisation during process development.
3P Instruments offers numerous possibilities, to ensure the integrity, functionality and stability of biotherapeutics.
Biotherapeutics are subject to stringent requirements in terms of safety, efficacy, reliability, integrity and functionality. During the production process, storage and transport, these requirements must be met without interruption in order to guarantee the patient consistency in the healing process. This is ensured by standardised quality controls from batch to batch. Only when all quality criteria for the structure and functionality of a batch have been met it is released for the pharmaceutical market. Even minor structural changes can have a negative impact on the safety and efficacy of biotherapeutics. Different analytical methods help with the batch characterisation of biotherapeutics.
3P Instruments offers numerous options for the quality control of biotherapeutics to ensure their safety and efficacy.
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