Molecular expression systems can be used to produce whole antibodies or antibody fragments. The properties of these expression products can be tested in assays of binding or pathogenicity. Expression systems can be used to produce large quantities of antibodies which are already well-characterized,to produce new antibodies by repertoire cloning, or to produce slight modifications in the sequences of antibodies by mutagenesis prior to expression. This paper reviews the ways in which these methods have been used to study the structure and function of human and murine anti-DNA antibodies. A consistent finding, from experiments using a range of different expression methods and antibodies, is that sequence motifs including arginine residues play a major role in binding to DNA. These motifs can be present on either the heavy or the light chain, but are particularly reported in VHCDR3.
Rahman A , Giles I , Haley J . Systematic analysis of sequences of anti-DNA antibodies— relevance to theories of origin and pathogenicity . Lupus2002; 12: 807–823 .
2.
Johnson KS , Chiswell DJ . Human antibody engineering . Curr Opin Struct Biol1993; 3: 564–571 .
3.
Barbas SM , Ditzel HJ , Salonen EM , Yang W-P , Silverman GJ , Burton DR . Human autoantibody recognition of DNA . Proc Natl Acad Sci USA1995; 92: 2529–2533 .
4.
Radic MZ , Weigert M . Genetic and structural evidence for antigen selection of anti-DNA antibodies . A Rev Immunol1994; 12: 487–520 .
5.
Rahman A , Latchman DS , Isenberg DA . Immunoglobulin variable region sequences of human monoclonal anti-DNA antibodies . Semin Arth Rheum1998; 21: 141–154 .
6.
Roben P , Barbas SM , Sandoval L et al.. Repertoire cloning of lupus anti-DNA autoantibodies . J Clin Invest1996; 98: 2827–2837 .
7.
Seal SN , Hoet RMA , Raats JMH , Radic MZ . Analysis of autoimmune bone marrow by antibody-phage display . Arthritis Rheum2000; 43: 2132–2138 .
8.
Brigido MM , Polymenis M , Stollar BD . Role of mouse VH10 and VLgene segments in the specific binding of antibody to Z-DNA, analyzed with recombinant single chain Fv molecules . J Immunol1993; 150: 469–479 .
9.
Polymenis M , Stollar BD . Critical binding site amino acids of anti-Z DNA single chain Fv molecules: role of heavy and light chain CDR3 and relationship to autoantibody activity . J Immunol1994; 152: 5318–5329 .
10.
Mockridge CI , Chapman CJ , Spellerberg MB , Isenberg DA , Stevenson FK . Use of phage surface expression to analyze regions of a human V4-34 (VH4-21)-encoded IgG autoantibody required for recognition of DNA: no involvement of the 9G4 idiotope . J Immunol1996; 157: 2449–2454 .
11.
Katsube Y , Suzuki H , Muryoi T , Sasaki T . Analysis of kappa light chain contribution to anti-DNA antibody activity of a human VH4-21-encoded monoclonal antibody (NE-1) by antibody-phage display technique . Int J Mol Med1998; 1: 863–868 .
12.
Lecerf J-M , Chen Y , Richalet-Secordel P , Wang X , Stollar BD . Autoreactivity of human VH domains from cDNA libraries: analysis with a bacterial expression system . J Immunol1998; 161: 1274–1283 .
13.
Kumar S , Kalsi J , Ravirajan CT et al.. Molecular cloning and expression of the Fabs of human auto-antibodies in Escherichia coli: determination of the heavy or light chain contribution to the anti-DNA/cardiolipin activity of the Fab . J Biol Chem2000; 275: 35129–35136 .
14.
Kumar S , Kalsi J , Latchman DS , Pearl LH , Isenberg DA . Expression of the Fabs of human auto-antibodies in Escherichia coli: optimization and determination of their fine binding characteristics and cross-reactivity . J Mol Biol2001; 308: 527–539 .
15.
Kalsi JK , Martin ACR , Hirabayashi Y et al.. Functional and modelling studies of the binding of human monoclonal anti-DNA antibodies to DNA . Mol Immunol1996; 33: 471–483 .
16.
Rahman A , Haley J , Radway-Bright E et al.. The importance of somatic mutations in the Vlgene 2a2 in human monoclonal anti-DNA antibodies . J Mol Biol2001; 307: 149–160 .
17.
Radic MZ , Mascelli MA , Erikson J , Shan H , Weigert M . Ig H and L chain contributions to autoimmune specificities . J Immunol1991; 146: 176–182 .
18.
Radic MZ , Mackle J , Erikson J , Mol C , Anderson WF , Weigert M . Residues that mediate DNA binding of autoimmune antibodies . J Immunol1993; 150: 4966–4977 .
19.
Stevenson FK , Longhurst C , Chapman CJ et al.. Utilization of the VH4-21 gene segment by anti-DNA antibodies from patients with SLE . J Autoimmun1993; 6: 809–826 .
20.
Katz JB , Limpanasithikul W , Diamond B . Mutational analysis of an autoantibody: differential binding and pathogenicity . J Exp Med1994; 180: 925–932 .
21.
Mostoslavsky G , Fischel R , Yachimovich N et al.. Lupus anti-DNA antibodies cross-react with a glomerular structural protein: a case for tissue injury by molecular mimicry . Eur J Immunol2001; 31: 1221–1227 .
22.
Van Bruggen MCJ , Walgreen B , Rijke TPM et al.. Antigen specificity of anti-nuclear antibodies complexed to nucleosomes determines glomerular basement membrane binding in vivo . Eur J Immunol1997; 27: 1564–1569 .
23.
Pewzner-Jung Y , Simon T , Eilat D . Structural elements controlling anti-DNA antibody affinity and their relationship to anti-phosphorylcholine activity . J Immunol1996; 156: 3065–3073 .
24.
Zack DJ , Yamamoto K , Wong AL , Stempnak M , French C , Weisbart RH . DNA mimics a self-protein that may be a target for some anti-DNA antibodies in systemic lupus erythematosus . J Immunol1995; 154: 1987–1994 .
25.
Haley JD , Rahman A , Latchman DS , Isenberg DA . Stable transfection of CHO cells to express whole human IgG anti-DNA antibodies . Lupus2001; 10(Suppl 1): S52-S52 .
26.
Li Z , SchettinoEW , PadlanEA , Ikematsu H , Casali P . Structure–function analysis of a lupusanti-DNA autoantibody:central role of the heavy chain complementarity determining region 3 arginine in binding of double and single stranded DNA . Eur J Immunol2000; 30: 2015–2026 .
