Synthetic Antigen Controls for Immunohistochemistry.

Area of Science:

• Biotechnology
• Histology
• Immunology

Background:

• Immunohistochemistry (IHC) relies on accurate positive and negative controls for interpreting protein expression.
• Availability of suitable tissue or cell line controls for IHC can be limited.

Purpose of the Study:

• To present a simple, adaptable protocol for generating synthetic immunohistochemistry (IHC) controls.
• To enable creation of controls for various antigen types and concentrations.

Main Methods:

• Synthetic controls were created using serial dilutions of a human erythroblastic oncogene B2 (ERBB2/HER2) intracellular domain (ICD) peptide in bovine serum albumin (BSA).
• The peptide/BSA mixture was cross-linked with formaldehyde and heated, then processed, sectioned, and stained similarly to tissue samples.

Main Results:

• The protocol yields a series of samples with known antigen concentrations, demonstrating a wide range of staining intensities.
• The method is compatible with routine histology laboratory procedures.

Conclusions:

• This technique provides a straightforward way to generate well-characterized synthetic IHC controls when endogenous controls are lacking.
• The ability to create controls with defined antigen levels aids in assessing assay performance, dynamic range, and optimization.