Beyond Optimization
Optimization and Validation
It is our experience that many published data rely on poorly characterized and functioning bio-tools (e.g., tissues, antibodies, methods). This may lead to incorrect claims regarding the distribution and regulation of a biomarker (representing a drug target, efficacy read-out or a pathological process)
The data variance introduced by differing qualities can also easily lead to complete loss of information regarding regulation of diagnostic or efficacy biomarkers
Going beyond the general optimisation procedures of many labs, Offspring have introduced a process that includes assay validation in addition to assay optimisation. To generate accurate data consistently and robustly it is critical to ensure antibodies and methodologies are fit-for-purpose
We optimise, validate and evaluate to demonstrate the processes of these investigative outcomes are consistent and accurate
Procurement of high-quality tissues (animal and human) is essential and ensured by adopting standardized handling, processing and storage procedures. Offspring have developed efficient methods to evaluate whether protein antigens are well preserved in the samples. This is achieved by IHC tissue-screening for carefully selected tissue antigens expressed in all cell types. This allows us to identify and remove tissue samples of inferior quality from the analyses and has particular importance when dealing with post-mortem tissues
By carefully selecting the antibodies and optimizing the various steps in IHC assay staining protocols as well as evaluating the tissue quality/integrity, we aim to achieve the likelihood that the data obtained are correct and reliable
In order to confirm IHC staining is real and specific for the target antigens of interest, Offspring have developed a series of validation procedures that will lead to the identification of specific staining (or not). Using molecular pathology techniques, ISH, proximity ligation assays, multiple control materials, image analysis and IHC we are able to demonstrate delivery of specific and consistently sensitive results
Establishment of Optimized and Validated assays
1. Identification and validation of one, and preferably two, high quality antibodies with pre-existing data that supports their usefulness for IHC analyses
2. Establishment of an optimized IHC staining protocol, using positive and negative control tissues
3. Employ the best performing antibody/IHC protocol to study tissues
4. Include pre-validated tissues in each experiment as historic controls
More detailed specifications for the Beyond workflow can be given on request and following execution of a mutual confidentiality disclosure agreement