Applications And Engineering Of Monoclonal Antibodies by David J. King

By David J. King

A worthwhile source for researchers and staff within the fields of either prescription drugs and biotechnology in addition to undergraduates in biochemistry, utilized biology, biomedical sciences and pharmacy, this booklet compares tested strategies of antibody construction with the recent. Antibody constitution and the consequences of antibody engineering are totally mentioned, and a case examine technique illustrates how antibodies are discovering expanding use within the prognosis and remedy of illness. the amount ends with advertisement expression, purification and large-scale manufacture of antibodies and their destiny strength, fairly as healing brokers.

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1996). Two different types of antibody fragment have been widely used for display of antibody specificities on phage, Fab and single-chain Fv (scFv, see Chapter 2). Antibody fragments are used as these are efficiently expressed in bacterial systems (see Chapter 5) and they can be well presented on the surface of the phage. , 1991). These phages are single-stranded DNA phages which consist of a long single strand of DNA coated with approx. 2700 copies of the major coat protein pVIII and a low number of copies of four minor protein components, including the pIII protein which is normally present at 3–5 copies.

This technology is very much in its infancy and, in the next few years, will no doubt develop to the point where a strain of mice can be produced which is capable of generating a range of useful human antibodies. 3, and many human MAbs have been generated, but the technology is far from routine and relatively few high affinity antibodies have been produced. Recently an alternative technology for the production of antibodies which can bypass hybridoma technology has been developed, known as phage display.

The source of lymphocytes is also a major problem. It is usually not possible to harvest these from the spleen or lymph node and the most available and commonly used source is peripheral blood. B cells are comparatively poorly represented in peripheral blood and the majority of those present express surface IgM. Fusion therefore often results in cells which largely produce low affinity IgM antibodies. The choice of cell line to be used as a fusion partner is also problematic. Human myeloma lines have proved to be comparatively poor with respect to their growth characteristics and fusion efficiencies and usually secrete immunoglobulin of their own which complicates production and purification.

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