SubscribeAs part of the biotechnological revolution, the first recombinant protein therapy - human insulin - was launched into the pharma market in 1982. Since then 119 additional recombinant proteins have been released – among them 6 blockbusters.
In the last five years, the growth rate of the US market for biopharmaceuticals has exceeded 28%. The growth rate of this particular pharma sector has doubled twice as fast as the classical pharma market.
While the young pharma products are certainly attractive, their quality control presents a challenge for the biopharma market. Based on a long period of experience, quality controls for classical small molecules have already been established; however, setting quality controls for protein therapeutics continues to be demanding. This is a result of the manufacturing process as well as the fact that the product it-self is complex and often heterogeneous.
ICH Guideline Q6B
Establishing a comprehensive characterization profile is a crucial step for recombinant protein quality control. Such a characterization profile allows for the setting of appropriate specifications. The ICH Guideline Q6B, Test Procedures and Acceptance Criteria for Biotechnological / Biological Products provides important information for the development of such profiles. These guidelines state that a characterization program should analyze physicochemical parameters, biological activity, content, and purity.
Physicochemical Parameters
Physicochemical parameters determine the protein's structure, physicochemical characteristics, and content. Information about the primary structure can be obtained from the combination of results from N- and C-terminal sequencing, amino acid analysis, and peptide mapping. Data about the secondary- and tertiary structure can be collected by spectroscopic methods; for example, by fluorescence or CD. Additionally, physicochemical protein characteristics can be obtained by the determination of the molecular weight, isoform patterns, and extinction coefficient. Determining the separation pattern can be achieved by electrophoretic or chromatographic methods. The separation pattern provides evidence regarding identity as well as data regarding homogeneity and purity.
Biological Activity
Assessing biological activity is another essential step in establishing a characterization profile. This parameter is defined as the capacity of a product to achieve a defined biological effect. The biological activity data provides functionality information and, therewith, indirectly provides information about proper protein folding.
Content
Typically, content is determined by an absorption measurement at 280 nm, a quantitative amino acid analysis, or a HPLC- or capillary electrophoretic method.
The rotor of an analytical ultracentrifuge
Purity
Impurities can be divided into two categories: product-related or process-related. Product- related impurities are product modifications such as oxidation or deamidation, aggregation or degradation. Process-related impurities stem from the cells, the cell culture, or the purification process, itself. These impurities include the DNA and protein of the host cell, the cell media components such as growth hormones, and substances that will be used in the down-stream process. These substances are utilized for cell lyses and protease inhibition as well as solubilization and virus inactivation. Additional impurities are substances from the chromatographic system, itself - the so-called "leachables" such as ligands for affinity chromatography.
Solvias is Your Preferred Partner for Protein Therapeutics Analysis - Experienced, Flexible, and Reliable
This wide range of testing for characterization profiling reflects the demand for a wide range of state of the art instrumentation, extensive expertise, and high quality standards. If such characterization programs were outsourced, the complex production process would force the analytics partner to meet two important requirements. First, the partner would have to be extremely flexible. Second, the partner would have to be highly reliable, guaranteeing on-time delivery for projects with a short turn-around time.
Solvias prides itself on its many years of intensive and extensive experience in protein therapeutic analytics, and is able to provide you with the highest degree of reliability. In its efforts to be-come even more flexible, Solvias has combined resources for this product area and raised the corresponding project manager level. Solvias wants to be your preferred partner for providing comprehensive characterization programs and for solving complex problems - such as analyzing carbohydrate structures or measuring detergents in the ppm range.
The separation of protein pI makers with CE-IEF