title: The role of in vitro expression systems in the investigation of antibodies to DNA.
abstract: OBJECTIVE
  Antibodies to DNA are believed to be important in the development of tissue inflammation and clinical activity in systemic lupus erythematosus (SLE). Sequence analysis of monoclonal murine and human anti-DNA antibodies suggests that somatic mutations and basic residues are important features at the DNA-binding site. To test this hypothesis, it is possible to alter these residues by site-directed mutagenesis of cloned variable region cDNA. The mutagenized cDNA sequence is then expressed in the form of a protein molecule whose properties can be tested in assays of binding or pathogenicity. The purpose of this article is to provide a systematic review of the evidence derived by such methods in the study of anti-DNA antibodies.
  
  
  METHODS
  Various different expression systems are available. Experiments using bacterial and eukaryotic expression systems are considered in turn. The advantages and disadvantages of the systems are described and the results obtained are compared.
  
  
  CONCLUSIONS
  High yields of antibody fragments such as scFv and Fab can be achieved by expression in bacteria. Such studies tend to confirm that reversion of somatic mutations or removal of basic residues at the antigen binding site reduce affinity for DNA. Tests of pathogenicity can only be performed by expressing whole antibodies in eukaryotic cells. The limited data available from expression of mutagenized cDNA in such systems argue against a simple relationship between changes in DNA binding affinity and changes in pathogenic potential. Further studies are therefore required to analyze the sequence requirements for pathogenicity.
authors.affiliation: Department of Medicine, University College, London, UK.
authors.forename: A
authors.initials: A
authors.lastname: Rahman
authors.forename: D S
authors.initials: DS
authors.lastname: Latchman
authors.forename: D A
authors.initials: DA
authors.lastname: Isenberg
mesh_terms: D000818:Animals; D000974:Antibodies, Antinuclear / Q000235:genetics* / Q000276:immunology*; D015870:Gene Expression / Q000276:immunology; D006801:Humans; D008180:Lupus Erythematosus, Systemic / Q000235:genetics* / Q000276:immunology*
chemical_list: D000974:Antibodies, Antinuclear