Related Experiment Video
Updated: Sep 25, 2025

Demonstration of the Sequence Alignment to Predict Across Species Susceptibility Tool for Rapid Assessment of Protein Conservation
Published on: February 10, 2023
Too Sensitive or Just Right?
Dana Martin1, Steven C Smith1,2, Alden Chesney1
1Department of Pathology, Virginia Commonwealth University (VCU) School of Medicine, Richmond, VA, USA.
The D5F3 antibody clone demonstrated superior performance in detecting anaplastic large cell lymphoma (ALCL) compared to the ALK1 clone, showing greater staining intensity and proportion. This finding is crucial for accurate ALCL diagnosis, especially with limited tissue samples.
Area of Science:
- Oncology
- Immunohistochemistry
- Molecular Pathology
Background:
- Anaplastic large cell lymphoma (ALCL) diagnosis relies on accurate assessment of anaplastic lymphoma kinase (ALK) expression.
- Established immunohistochemical markers, such as the ALK1 clone, are used for ALK assessment in ALCL.
- Novel antibody clones are continuously evaluated to improve diagnostic sensitivity and specificity.
Purpose of the Study:
- To compare the diagnostic performance of a novel rabbit monoclonal antihuman CD246 antibody (D5F3 clone) against the ALK1 clone for ALCL.
- To evaluate the intensity and proportion of neoplastic cells positive for ALK expression using both antibody clones.
Main Methods:
- Archival ALCL cases (n=27) underwent immunohistochemical assessment using ALK1 and D5F3 clones.
- Staining intensity and proportion of positive neoplastic cells were evaluated using a standardized scoring system.
- Results were compared with clinical anaplastic lymphoma kinase (ALK) break-apart fluorescence in situ hybridization (FISH) assays.
Main Results:
- D5F3 clone identified more ALK-positive ALCL cases (48%) compared to ALK1 clone (33%).
- D5F3 staining demonstrated significantly greater proportion (P=.02) and intensity (P=.03) of positive cells.
- Three cases positive for D5F3 but negative for ALK rearrangements by FISH showed copy number gains at the ALK locus in a subset of cells.
Conclusions:
- The D5F3 clone exhibits enhanced sensitivity and intensity for ALK detection in ALCL compared to the ALK1 clone.
- The D5F3 clone's superior performance is particularly beneficial in cases with limited tissue samples.
- Orthogonal ALK testing methods are recommended for cases with weak or focal staining to ensure diagnostic accuracy.
More Related Videos
09:39Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications
Published on: January 27, 2014
12:04Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-cut Lung Slices
Published on: May 9, 2018
Related Concept Videos
Sensitivity, Specificity, and Predicted Value
Sensitivity is the...
Decision Making: Traditional Method
First, a specific claim about the population parameter is decided based on the research question and is stated in a simple form. Further, an opposing statement to this claim is also stated. These statements can act as null and alternative hypotheses, out of which a null hypothesis would be a...
Assumptions of Survival Analysis
Statistical Analysis: Overview
One of the most commonly used statistical quantifiers is the mean, which is the ratio between the sum of the numerical values of all results and the...
Types of Biopharmaceutical Studies: Controlled and Non-Controlled Approaches
Non-controlled studies, commonly employed for initial exploration, lack a control group, rendering them susceptible to biases and external influences. In contrast,...
Significance Testing: Overview