Conjugation of acetylcholine receptor-protecting Fab fragments with polyethylene glycol results in a prolonged half-life in the circulation and reduced immunogenicity.
Antibodies to the acetylcholine receptor (AChR) cause AChR loss, resulting in the disease, myasthenia gravis (MG). The majority of the pathogenic antibodies seem to be directed against the main immunogenic region (MIR) of the AChR. In contrast to the intact antibodies, Fab fragments of anti-AChR antibodies are not themselves pathogenic and such fragments of anti-MIR monoclonal antibodies (mAbs) protect the AChR in vitro and in vivo against the pathogenic antibodies. However, Fab fragments have a very short in vivo half-life and are immunogenic, obstacles which must be overcome before their clinical use can be envisaged. We investigated the effect of conjugating Fab fragments to polyethylene glycol (PEG), a method known to increase the in vivo half-life and reduce the immunogenicity of proteins. When the Fab' fragments of two rat anti-MIR mAbs (nos. 35 and 195) were conjugated to methoxy-PEG-maleimide, the conjugates retained about 10% of their AChR binding activity and efficiently protected the AChR against the binding and modulating activity of myasthenic antibodies. Their in vivo half-life in rats was approximately 15 times longer than that of the unconjugated Fab' fragment and they were much less immunogenic in mice. This work represents an important step towards the clinical use of AChR-protective anti-MIR Fabs, but further improvements are needed before their clinical use is attempted.
Trakas
N
N
Department of Biochemistry, Hellenic Pasteur Institute, 127 Vas. Sofias Avenue, Athens 115 21, Greece.
Tzartos
S J
SJ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001323:Autoantibodies / Q000737:chemistry* / Q000276:immunology; D001667:Binding, Competitive / Q000187:drug effects / Q000276:immunology; D006207:Half-Life; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009157:Myasthenia Gravis / Q000188:drug therapy* / Q000276:immunology / Q000503:physiopathology; D018696:Neuroprotective Agents / Q000138:chemical synthesis* / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D007140:Immunoglobulin Fab Fragments; D018696:Neuroprotective Agents; D011950:Receptors, Cholinergic; D014614:Vaccines, Synthetic; D011092:Polyethylene Glycols