Although the development of hybridoma technology (see Chapter 1) was a critical breakthrough in life science research, the technology has a number of limitations. It requires considerable time, expense, and expertise, as well as specialized cell culture facilities, and relies on the use of animals. Additionally, many molecules are not immunogenic in mammals or are toxic and cannot be used as antigens. Most seriously for therapeutic application of antibodies, standard methods of producing monoclonal antibodies (MAbs) yield rodent antibodies that are rejected by the human immune system. Various approaches to overcome this limitation have been attempted, such as combining the DNA that encodes the binding site of mouse MAbs with DNA that encodes human antibodies. This generates antibodies known as either chimeric antibodies or humanized antibodies, depending on how large a part of the mouse antibody is used. Another approach is to use mice that are genetically engineered to produce antibodies with human antibody sequences.

From: Methods in Molecular Biology, vol. 378: Monoclonal Antibodies: Methods and Protocols Edited by: M. Albitar © Humana Press Inc., Totowa, NJ

Developments in the fields of bacterial expression of functional antibodies and methods to select genes from a library by using the phenotype of the encoded polypeptide between 1985 and 1988 have been a breakthrough in antibody technology.

This chapter summarizes some of the methodologies commonly used to select, identify, and engineer antibody fragments for particular properties and apply them in typical immunoassays.

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