The objective of bioassays is to determine the biological activity and to analyze the mode of action of biological products as part of GMP regulations.
What are the GMP requirements of bioactivity testing?
About Bioassay Validation?
Bioassays play an important role in the product characterization of biological products. The assessment of the biological activity (potency) is part of the regulatory GMP requirements as described in the quality guideline ICH Q6B.
About Potency Assays
The main objective of a potency assay is to determine the relative potency of GMP release of samples.
Bioassay Validation in compliance with United States Pharmacopeia (USP) <1033>
The objective of bioassay validation is to verify that the assay is fit for the intended purpose.
A validated bioassay characterizes the performance of the assay and its limitations, provides confidence in data quality and contributes to GMP compliance for batch release and submission for manufacturing authorization and clinical trial studies.
The reason for performing the assay must be distinctly specified, e.g. batch release, stability, comparability, ect. The GMP requirements are stated in the ICH Q6B Quality Guideline, the validation guidelines are stated in the US Pharmcopeia (USP).
Bioactivity Testing
The assessment of the biological activity is part of the characterization profile as described in ICH Q6B. The bioactivity is depending on the overall shape of the protein which is laid out by the amino acid sequence.
How the protein is folded determines its function. Subtle changes in the protein structure can alter its bioactivity. Glycosylation is one of the most commonly known post-translation modifications.
Consistency in assays is mandatory for validation experiments and statistical analysis. Therefore, skilled, experienced and trained technicians are essential and challenging within your work environment.
Principles and Practices with Statistical Analysis
Dose-response curve
In some bioassays, a linear response model can be applied, for statistical analysis, to quantitatively determine the analyte. However, the data type of the bioassay model to analyze the cellular response, follows in most cases a sigmoidal dose-response curve. The dose-response curve represents a graphical illustration of the analyte (x-values) and the biological effect (y-values).
Four parameter logistic regression model
The four-parameter logistic regression model is quite useful for the statistical analysis of cell-based assays for dose-response analysis. It is a sigmoidal-shaped, nonlinear regression model used to quantify the analyte. Four parameters (minimum asymptote, maximum asymptote, inflection point (EC50), and Hill coefficient) define the validation characteristics of the test performance in which the inflection point reflects the EC50 value.
EC50
The EC50 value defines the half-maximal effective concentration of the analyte. The EC50 value is used to determine the potency of proteins.
Hands-on Experience
- Proficiency with cell and molecular biology
- Proficiency with Florescence Activated Cell Sorting (FACS)
- Proficiency with immunofluorescence analysis
- Proficiency with cell-based Assay procedures and Potency Assays
- Proficiency with mammalian cell culture
- Proficiency with regulatory GMP requirements (FDA, EMA, EU)
- Proficiency with Specifications for Biological Products according to ICH Q6B
On-site Services
- Potency Assays
- Cell-based assays for Drug Screening and Target Validation
- Statistical Analysis of Validation Experiments
Key Benefits
- Contributing to validated bioassays
- Contributing to GMP release of samples
- Contributing to clinical trial and marketing authorization
