Preparation of an immunoadsorbent coupled with a recombinant antigen to remove anti-acetylcholine receptor antibodies in abnormal serum.
An immunoadsorbent that removes anti-acetylcholine receptor antibodies (AChRAb) in abnormal serum of myasthenia gravis (MG) patient was efficiently prepared by an expression product, the functional fragment of AChR(alpha205) fused with maltose binding protein (MBP). The ligand can then covalently bind to amylose resin through MBP fusion protein. It was shown from the result of this study with anti-AChR mice sera that the removal rate of AChRAb on this immunoadsorbent reached 87+/-10% (mean value of 10 mice) and the maximally binding capacity of AChRAb was approximately 260 microg/g immunoadsorbent (wet weight). Moreover, the immunoadsorption test of sera in two MG patients indicated that about 90% and 96% of abnormal AChRAb could be eliminated, while other serum components such as albumin, IgG, IgM and IgA only dropped 18%, 35%, 22%, 15% and 24%, 27%, 15%, 12%, respectively, for two MG patient sera. It is anticipated from this study that the immunoadsorbent reported here could, with further development, find its clinical application for removal of AChRAb from patient serum.
Guo
Chen-Yun
CY
Institute of Biotechnology, Laboratory of Chemical Biology and Molecular Engineering of National Education Ministry, Shanxi University, 580 Wucheng Road, Taiyuan 030006, PR China.
Li
Zhuo-Yu
ZY
Xu
Ming-Qun
MQ
Yuan
Jing-Ming
JM
D000818:Animals; D001323:Autoantibodies / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism; D001324:Autoantigens / Q000235:genetics* / Q000276:immunology; D001666:Binding Sites, Antibody; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D007163:Immunosorbent Techniques; D007164:Immunosorbents / Q000138:chemical synthesis* / Q000378:metabolism*; D058619:Maltose-Binding Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D009157:Myasthenia Gravis / Q000097:blood / Q000276:immunology; D010957:Plasmids; D011950:Receptors, Cholinergic / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis* / Q000378:metabolism
D001323:Autoantibodies; D001324:Autoantigens; D002352:Carrier Proteins; D007164:Immunosorbents; D058619:Maltose-Binding Proteins; D011950:Receptors, Cholinergic; D011993:Recombinant Fusion Proteins