Antibody Synthesis In Vitro


Antibodies are specific proteins that recognise nearly any chemical structure. During in vivo immunisation B cells are activated by antigen. Antigen‐specific cells proliferate and some differentiate into antibody‐secreting plasma cells. Monoclonal antibody technology was first reported by Köhler and Milstein in 1975. Briefly, B cells with specificity for a known antigen were fused with myeloma to make them immortal. Hybridomas grow in culture indefinitely and secrete monoclonal antibodies (mAbs) of a desired specificity. Hybridoma technology and antibody engineering have contributed in Biomedicine to the diagnosis of malignancies, tissue typing, identification of individual cell types, detection of cytokines, growth factors and hormones. The mAbs have become important in human immunotherapy and diagnosis. Murine mAbs have limited potential in human therapeutic applications due to human immune response to the murine antibodies. Several strategies for human mAbs production for immunotherapy are described in this article. Monoclonal antibodies have also a wide range of applications in Health and Food Sciences, Environmental Sciences, Technology and Experimental Sciences among others.

Key Concepts

  • Antigen is a molecule that binds specifically to B cell receptor (BCR) or soluble antibodies. T cell receptor (TCR) recognise processed antigen presented by MHC (major histocompatibility complex).
  • Epitope or antigenic determinant is part of an antigen that is recognised specifically by TCR‐MHC or by BCR or by soluble antibodies.
  • Immunogen is an antigen that generates a cellular and humoral immune response.
  • Antibody is an immunoglobulin that recognises specifically a particular antigenic epitope.
  • Antibody affinity is the strength of binding between a single binding site of an antibody and a single antigenic determinant or epitope.
  • Plasma B cells are differentiated B cells that, after binding to an antigen, secrete soluble antibodies.
  • Hybridoma is a hybrid cell line obtained by cell fusion of an immortalised lymphocyte tumor line, or myeloma, and B lymphocytes with defined antigen specificities.
  • Monoclonal antibody is an immunoglobulin with specificity for one antigenic determinant or epitope, derived from a single B‐clone.
  • Antibody immunotherapy is applied in immunopathology and in cancer. The main therapeutic agents are human or humanised monoclonal antibodies.

Keywords: specific antibodies; monoclonal antibody technology; hybridomas; B cell diversity; B cell immortalisation

Figure 1. Generation of monoclonal antibody secretory hybridomas by B cell fusion. (a) In vivo, high‐affinity B cells are developed in germinal centres and plasma B cells secrete high‐affinity antibodies.
Figure 2. Several strategies for human monoclonal antibody production. (a) human–mouse heterohybridoma, (b) antibody‐transfected cell line, (c) virus immortalised B cells, (d) human–human hybridoma, (e) transgenic mouse hybridoma.


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Iborra, Antoni, Martínez, Paz, and Palacio, José Ramón(Jul 2015) Antibody Synthesis In Vitro . In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0001115.pub2]