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Review
. 2000 Jun;53(3):111-7.
doi: 10.1136/mp.53.3.111.

Monoclonal antibodies

P N Nelson  1 , G M Reynolds, E E Waldron, E Ward, K Giannopoulos, P G Murray
Affiliations
Review

Monoclonal antibodies

P N Nelson et al. Mol Pathol. 2000 Jun.

Abstract

Monoclonal antibodies are essential tools for many molecular immunology investigations. In particular, when used in combination with techniques such as epitope mapping and molecular modelling, monoclonal antibodies enable the antigenic profiling and visualisation of macromolecular surfaces. In addition, monoclonal antibodies have become key components in a vast array of clinical laboratory diagnostic tests. Their wide application in detecting and identifying serum analytes, cell markers, and pathogenic agents has largely arisen through the exquisite specificity of these unique reagents. Furthermore, the continuous culture of hybridoma cells that produce these antibodies offers the potential of an unlimited supply of reagent. In essence, when compared with the rather limited supply of polyclonal antibody reagents, the feature of a continuous supply enables the standardisation of both the reagent and the assay technique. Clearly, polyclonal and monoclonal antibodies have their advantages and disadvantages in terms of generation, cost, and overall applications. Ultimately, monoclonal antibodies are only produced when necessary because their production is time consuming and frustrating, although greatly rewarding (at least most of the time!). This is especially apparent when a monoclonal antibody can be applied successfully in a routine pathology laboratory or can aid in the clinical diagnosis and treatment of patients. In this article, the generation and application of monoclonal antibodies are demystified to enable greater understanding and hopefully formulate novel ideas for clinicians and scientists alike.

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Figures

Figure 1
Figure 1
Schematic representation of an antibody molecule highlighting the “Y” shaped structure.
Figure 2
Figure 2
Five stages of generating a murine monoclonal antibody (MAb). (A) Immunisation, illustrating tail bleeds from mice immunised with Epstein-Barr virus (EBV) latent membrane protein 1 multiple antigenic synthetic peptide. (B) Fusion and selection, showing hybridoma PNG312G. (C) Screening, highlighting reactivity of MAb PNG211D against human osteoclastoma. (D) Characterisation, epitope mapping of MAb A57H against human IgG Fc. (E) Further developments, molecular modelling of monoclonal antibody A57H revealing an epitope (red) in the CH3 domain of IgG. CFA, complete Freund's adjuvant; ELISA, enzyme linked immunosorbent assay; HA, haemagglutination; HAT, hypoxanthine, aminopterin, and thymidine; HT, hypoxanthine and thymidine; IC, immunochemistry; IFA, incomplete Freund's adjuvant; PEG, polyethylene glycol; WBLOT, western blotting.
Figure 3
Figure 3
A monoclonal antibody directed against cytokeratin demonstrates micrometastases in smooth muscle of large bowel.
Figure 4
Figure 4
The use of a monoclonal antibody in the immunocytochemical detection of high HER-2/neu expression in a metastatic deposit of breast cancer.
See this image and copyright information in PMC

References

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    1. Hudson PJ. Recombinant antibody constructs in cancer therapy. Curr Opin Immunol 1999;11:548–57. - PubMed

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