Related Experiment Video
Updated: Jun 27, 2026
Synthetic Antigen Controls for Immunohistochemistry
Published on: August 23, 2021
Experimental validation of peptide immunohistochemistry controls
Steven A Bogen1, Kodela Vani, Brian McGraw
1Medical Discovery Partners LLC, USA. sbogen@bu.edu
Insights
New peptide immunohistochemistry (IHC) controls offer a reliable method for quality assurance in clinical laboratories. These controls accurately reflect IHC staining performance, enhancing diagnostic accuracy and standardization in automated manufacturing.
Area of Science:
- Biotechnology
- Clinical Diagnostics
- Quality Control
Background:
- Peptide immunohistochemistry (IHC) controls represent a novel format for verifying IHC assay performance.
- Formalin-fixed peptide epitopes on slides mimic native protein behavior in tissue sections.
- Existing quality control methods may not fully capture the complexities of IHC assays in diverse clinical settings.
Purpose of the Study:
- To evaluate the hypothesis that peptide IHC controls accurately reflect IHC staining performance in clinical laboratories.
- To assess the utility of peptide IHC controls as a practical quality assurance tool for clinical diagnostics.
- To determine the reproducibility and stability of the developed peptide IHC controls.
Main Methods:
- Development of an instrument for reproducible printing of peptide controls on microscope slides.
- Creation of software for automated measurement of stained control color intensity.
- A national study involving 109 clinical laboratories to correlate peptide control staining with HER-2 tissue staining.
Main Results:
- Automated printing of peptide spots demonstrated high reproducibility (4-8% CV).
- Peptide controls exhibited stability at <=4°C for at least 7 months.
- A strong correlation (r=0.87) was observed between peptide control staining and 3+ HER-2 tissue staining across laboratories.
Conclusions:
- Peptide IHC controls accurately reflect the analytical performance of IHC staining, including antigen retrieval.
- These controls are suitable for proficiency testing and verifying intralaboratory staining consistency using Levy-Jennings charts.
- Peptide IHC controls offer a standardized and high-throughput compatible solution for IHC quality assurance.
Abstract:
Peptide immunohistochemistry (IHC) controls are a new quality control format for verifying proper IHC assay performance, offering advantages in high throughput automated manufacture and standardization. We previously demonstrated that formalin-fixed peptide epitopes, covalently attached to glass microscope slides, behaved (immunochemically) in a similar fashion to the native protein in tissue sections. To convert this promising idea into a practical clinical laboratory quality control tool, we tested the hypothesis that the quality assurance information provided by peptide IHC controls accurately reflects IHC staining performance among a diverse group of clinical laboratories. To test the hypothesis, we first designed and built an instrument for reproducibly printing the controls on microscope slides and a simple software program to measure the color intensity of stained controls. Automated printing of peptide spots was reproducible, with coefficients of variation of 4% to 8%. Moreover, the peptide controls were stable at
