Experimental validation of peptide immunohistochemistry controls

• Published in: Applied immunohistochemistry & molecular morphology Vol. 17; no. 3; p. 239
• Main Authors: Bogen, Steven A, Vani, Kodela, McGraw, Brian, Federico, Vin, Habib, Iqbal, Zeheb, Ron, Luther, Ed, Tristram, Colin, Sompuram, Seshi R
• Format: Journal Article
• Language: English
• Published: United States 01.05.2009
• Subjects: Breast Neoplasms - diagnosis; Cell Line, Tumor; Epitopes - analysis; Epitopes - immunology; Humans; Immunohistochemistry - instrumentation; Immunohistochemistry - standards; Peptides - analysis; Peptides - immunology; Quality Control; Receptor, ErbB-2 - analysis; Receptor, ErbB-2 - immunology; Reference Standards; Software; Staining and Labeling - standards
• ISSN: 1533-4058
• DOI: 10.1097/PAI.0b013e3181904379

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 <or=4 degrees C for at least 7 months, the longest time duration we tested. A national study of 109 participating clinical laboratories demonstrated a good correlation between a laboratory's ability to properly stain formalin-fixed peptide controls to their ability in properly staining a 3+ HER-2 formalin-fixed tissue section mounted on the same slide (r=0.87). Therefore, peptide IHC controls accurately reflect the analytical component of an IHC stain, including antigen retrieval. Besides its use in proficiency survey testing, we also demonstrate the feasibility of applying peptide IHC controls for verifying intralaboratory IHC staining consistency, using Levy-Jennings charting.