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Molecular expression systems for anti-DNA antibodies—2

Institute of Cancer Research, Chester Beatty Labs, London, UK; Centre for Rheumatology, Bloomsbury Rheumatology Unit, Department of Medicine, University College London Hospital, Arthur Stanley House, 40–50 Tottenham Street, London W1P 9PG, UK; Antisoma Research Laboratory, St. Georges Hospital Medical School, Cranmer Terrace, London SW17 0QS, UK sanjeev.kumar{at}antisoma.com

J K Kalsi

C T Ravirajan

Centre for Rheumatology, Bloomsbury Rheumatology Unit, Department of Medicine, University College London Hospital, London, UK

D S Latchman

Institute of Child Health, London, UK

L H Pearl

Institute of Cancer Research, Chester Beatty Labs, London, UK

D A Isenberg

Centre for Rheumatology, Bloomsbury Rheumatology Unit, Department of Medicine, University College London Hospital, London, UK

Antibodies to double-stranded DNA are the best-known serological markers of systemic lupus erythematosus, and are closely associated with its renal pathogenesis. How these antibodies recognize DNA is not fully understood.An understandingof the relationship between the functional attributes of an antibody with the three-dimensional structure of its antigen-combining site would allow an insight into the rules that dictate auto-antibody–nucleic acid interaction and consequent pathogenicity of the autoantibody. Data from such studies could assist the development of novel drugs as an approach to specific therapies that can inhibit or disrupt protein–nucleic acid interactions. A full understanding of the binding specificities can be achieved only by experimental determination of detailed three-dimensional structure of these antibodies alone, and of their complexes with specific DNA antigens. A prerequisite of such a study is the ability to produce multimilligram quantities of the antibody protein. However, these antibodies are particularly difficult to express, probably due to their DNA-binding activity. This review attempts to focus on the recent developmentson the over-expressionof anti-DNA antibody fragments in heterologous cell expression systems and their purification to homogeneity that would in turn allow their structural studies via crystallization.

Key Words: human anti-DNA antibodies • heterologous cell expression systems • cloning • over-expression • purification

Lupus, Vol. 11, No. 12, 833-842 (2002)
DOI: 10.1191/0961203302lu304rr


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