Molecular pathology services

Biomarker method development

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 for 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 and Bond Rx III platforms, in most cases an entire IHC method can be developed and validated at Propath within as little as 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:

  • ‌Optimal positive and negative controls
  • ‌Heat induced antigen retrieval parameters, including enzyme retrieval
  • ‌The precision, specificity, linearity and reproducibility of the final method
  • ‌Selection of detection chemistry
  • ‌Strategies to amplify the signal and reduce non-specific background staining

Method validation report

‌Following optimisation and validation, full details of the method and validation results are provided to sponsors within a comprehensive validation report.

Propath’s reports include representative images of staining and all parameters required to reproduce staining. Whole slide digital scans can also be provided, demonstrating optimum working conditions in positive control tissue.

Reports can be audited for GLP and GCP compliance, where required.

Outsourced R&D - Done better.

At Propath we approach biopharma R&D differently. Say goodbye to Large CRO frustrations –and hello to a more responsive, agile and efficient level of service.