Garbage in, garbage out: A critical evaluation of strategies used for validation of immunohistochemical biomarkers

Abstract

The use of immunohistochemistry (IHC) in clinical cohorts is of paramount importance in determining the utility of a biomarker in clinical practice. A major bottleneck in translating a biomarker from bench‐to‐bedside is the lack of well characterized, specific antibodies suitable for IHC. Despite the widespread use of IHC as a biomarker validation tool, no universally accepted standardization guidelines have been developed to determine the applicability of particular antibodies for IHC prior to its use. In this review, we discuss the technical challenges faced by the use of immunohistochemical biomarkers and rigorously explore classical and emerging antibody validation technologies. Based on our review of these technologies, we provide strict criteria for the pragmatic validation of antibodies for use in immunohistochemical assays.

Commercialized biomarkers: new horizons in prostate cancer diagnostics

“…One key issue with tests based on IHC is antibody specificity. This issue can be overcome by establishing strict criteria for the pragmatic validation of antibodies for use in immunohistochemical assays as recently reviewed [68]. Antibody specificity is also common to ELISA assays which would otherwise have the advantage of being cheap, simple and reproducible.…”

Whole Slide Imaging for Analytical Anatomic Pathology and Telepathology: Practical Applications Today, Promises, and Perils

“…Developing techniques such as immunohistochemistry (IHC) and molecular diagnostics/theranostics have brought up the necessity and ability to measure discrete molecules or ''biomarkers'' within tissue; however, there are a number of analytical variables that can affect the results of these tests, including the accurate measurement of signals. 3,4 Given this inherent variability, there have been efforts to standardize these tests, and the US Food and Drug Administration, American Society of Clinical Oncology, and College of American Pathologists (CAP) have each provided special direction on some testing (eg, human epidermal growth factor 2 [HER2/neu]). 5,6 Whole slide imaging (WSI) with algorithmic analysis is one potential technology to improve the assessment of such markers (as shown in the Table).…”

Razvoj metoda imunohistokemijskog bojenja i optimizacija protokola za detekciju transkripcijskog faktora BACH2

“…Nakon odgovarajuće fiksacije, tkivo se dalje obrađuje da bi se moglo prožeti tekućim parafinom i pohraniti u obliku parafinskih blokova, koji se nakon toga mogu rezati u tanke rezove (1 do 5 µm) na mikrotomu. Tanki se rezovi potom prenose na adhezivna predmetna stakla na kojima su učvršćeni za daljnju obradu 2 …”