Developing a robust IHC method: Propath’s approach
Our objective when developing a novel protocol for immunohistochemistry, is to identify a method which produces highly specific staining with good signal intensity, where positive cells are clearly distinguishable from any negative cells or background staining. In addition, to be considered robust, a method should produce staining which is consistent and reproducible.
The team at Propath is experienced at developing, optimising and validating methods for biomarkers and test articles with no prior IHC staining information.
In addition to offering clients our own extensive portfolio of previously validated biomarker methods, we are able to develop and validate methods for most proprietary antibody structures, including antibody fragments, bi-specific antibodies, and antibody-drug conjugates (ADCs).
Using the very versatile Roche Ventana Discovery Ultra platform, in most cases an entire IHC method can be developed and validated at Propath within two weeks, although more complex antibody structures may take longer.
Features assessed during method development
Propath takes a detailed, systematic approach to method development, adjusting one single parameter at a time and evaluating the outcomes of that adjustment.
Propath’s standard approach for IHC method development and validation includes assessment of:
Method validation report
Following optimisation and validation, full details of the method are provided to sponsors within a comprehensive validation report, including representative images of staining where appropriate. Whole slide digital scans can also be provided, demonstrating optimum working conditions in positive control tissue.
Tissue Micro Arrays
Further testing of IHC protocols can be performed by Propath in TMAs, which provides additional insight into how an IHC protocol is likely to perform in a clinical trial involving multiple donors. In such cases, it is common for tissues to have undergone varying fixation and processing times, especially when they originate from multiple sites.