Monoclonal Antibodies: Diagnostic Uses

Abstract

Diagnosis of disease is a complex process requiring the clinician to deduce, from a set of symptoms and observations, an underlying cause, to predict which of the several therapeutic options is most likely to be effective, and to monitor that effectiveness. Laboratory tests that identify infectious agents, abnormal cells or elements of the body's response to disease, that quantify or localise particular molecules, cells or organisms in body fluids or tissues can provide valuable information to help the clinician with the initial diagnosis, with differential diagnosis (when the symptoms are consistent with several alternative causes) and with selection and monitoring of therapy. Antibodies, because of their exquisite specificity, are particularly useful reagents in this context, and monoclonal antibodies generally show superior specificity compared with polyclonal mixtures of antibodies. Monoclonal antibodies are used widely in the diagnosis of disease, whether in the diagnostic laboratory, in the doctor's rooms or in the field.

Key Concepts:

  • Correct diagnosis is a prerequisite to appropriate treatment of disease.

  • Antibodies make useful reagents for identifying infectious or other disease‐causing agents.

  • Monoclonal antibodies are particularly specific and often provide the best diagnostic reagents.

  • Antibodies can be used in a variety of assay formats, designed to answer different questions – presence or absence of a particular substance, amount present and localisation within tissues.

  • The rapid identification of new or rare infectious agents is an important public health measure, to monitor and reduce the chances of epidemics (as we see every few years with a new threat from an influenza variant).

  • Disease often results from imbalances or defects in normal physiological mechanisms, especially the immune system. Many diagnostic tests therefore analyse the components of normal physiological processes.

  • Laboratory tests are useful after the initial diagnosis, to monitor the effects of therapy.

  • Diagnostic tests are generally done in a specialised diagnostic laboratory, but it is often advantageous to perform tests at ‘point of care’ or at home.

Keywords: antibody; monoclonal; diagnosis; pathology; immunopathology

Figure 1.

Schematic representation of an ELISA for the detection and titration of IgG and IgM antibodies against the dengue virus. Courtesy of Peter Devine, Jody Mitchell and Andrea Cuzzubo, PanBio Pty Ltd.

Figure 2.

Typical application of monoclonal antibody in the differential diagnosis of leukaemia. Patterns of fluorescence intensity following FMC‐7 staining in three different disease states and in a healthy individual. (a) B‐cell chronic lymphocytic leukaemia (B‐CLL). FMC‐7 expression weakly positive. Clinical features: a high lymphocyte count, lymphoadenopathy and nonaggressive clinical course. (b) Prolymphocytic leukaemia (PLL). FMC‐7 expression strongly positive. Clinical features: enlarged spleen, high lymphocyte count, no lymphadenopathy and aggressive clinical course. (c) Hairy cell leukaemia (HCL). FMC‐7 expression strongly positive. Clinical features: enlarged spleen, marrow fibrosis, low white cell count and lymphadenopathy. (d) Healthy individual. FMC‐7 expression moderately positive on a normal B‐cell population. Courtesy of Dean Moss and Sean Meehan, AMRAD Biotech Pty Ltd.

Figure 3.

Detection of rhesus D‐positive fetal cells in maternal blood. The main panels show three maternal blood samples stained with antibody against the Rhesus D antigen. These mothers are Rhesus‐negative, but the fetuses have inherited Rhesus‐D positivity from their father. These very rare fetal cells in the maternal circulation can be detected using a suitable monoclonal antibody and flow cytometry. The inset over the left‐hand panel shows dual light‐scatter parameters used to select red cells for the analysis (indicated in red). Courtesy of Dean Moss and Sean Meehan, AMRAD Biotech Pty Ltd.

Figure 4.

A schematic diagram of cytometric bead array to detect the different analytes in the same sample using antibody‐coated capture beads with different fluorescence intensities. Diagram based on BD Biosciences CBA.

Figure 5.

Immunohistological analysis of needle biopsy from prostate tissue. The lobules of prostatic acini are normally surrounded by a layer of basal cells, which can be stained with an antibody against a high‐molecular weight cytokeratin and also for nuclear p63, a homologue of the tumour suppressor p53. In prostate cancer, this layer is generally absent. Though either marker is characteristic of these basal layer cells, the use of antibodies against both, detected with the same colour (brown) adds confidence. Alpha‐methylacyl‐CoA‐racemase (AMACR) is a protein that is greatly elevated in malignant prostate cells. This is detected with an antibody which is in turn revealed with a red colour. The combination of three antibodies, two which identify normal tissues and one which is greatly elevated in malignant tissues, increases the certainty with which prostate cancer can be identified and distinguished from nonmalignant conditions particularly in prostate needle biopsies containing a small amount of tumour tissue. The figure shows normal ducts (with brown staining) and malignant ducts (lacking brown staining, but heavily stained red) in the same tissue. Courtesy Dr Andrew Ruskiewicz, based on Tacha and Miller .

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Zola, Heddy, Mohandas, Arunesh Pullaniparambil, and Krumbiegel, Doreen(May 2013) Monoclonal Antibodies: Diagnostic Uses. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002177.pub3]