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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Xing, L. Zhang, G. H. Sun, L. Pan, S. Y. |
| Description | Country affiliation: China Author Affiliation: Sun L ( Department of Neurology, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, China.); Xing L ( Department of Neurology, The China Ordnance Northern Heavy Group Hospital (The Third Affiliated Hospital of Baotou Medical College), Baotou, China.); Zhang GH ( Department of Neurology, The Affiliated Hospital of Inner Mongolia Medical University, Hohhot, China.); Pan SY ( Department of Neurology, Nanfang Hospital, Southern Medical University, Guangzhou, China.) |
| Abstract | It is expensive to induce experimental autoimmune myasthenia gravis (EAMG) by active immunity, and difficult to obtain natural acetylcholine receptor (AChR). We sought a new method of inducing EAMG by immunizing rats with artificially synthesized AChR. The AChR mRNA in TE671 cells was extracted and reverse transcribed. The inclusion body was purified and protein concentration was determined, and the EAMG animal model was used for induction. The serum was extracted from rat blood. The antibody titer was determined using enzyme-linked immunosorbant assay (ELISA). The concentration of decay accelerating factor (DAF) in the rat serum was determined by ELISA, and the metabolism of serum rDAF was determined by western blot. We evaluated the inhibition of rDAF by determining the 50% complement hemolysis unit in the rat serum. The extracellular domain (ECD) nucleotide sequence clone produced by polymerase chain reaction was completely consistent with that in the human gene bank; it was induced by isopropyl ß-D-1-thiogalactopyranoside to express the protein after insertion into vector pET16b. Sodium dodecyl sulfate polyacrylamide gel electrophoresis demonstrated that the inclusion body protein was the exact target. The ECD protein was able to bind with mAb35 after dialysis and renaturation, which demonstrated protein activity. The soluble ECD protein was used to immunize rats and obtain the EAMG models. The inhibitory effect of the complement was unsatisfactory owing to high decay rate after rDAF injection into the EAMG models. It is easy to induce the EAMG model by obtaining the AChRTE 1 subunit ECD protein using the substitution method. |
| e-ISSN | 16765680 |
| Journal | Genetics and Molecular Research |
| Issue Number | 3 |
| Volume Number | 14 |
| Language | English |
| Publisher | Fundação de Pesquisas Científicas de Ribeirão Preto |
| Publisher Date | 2015-07-14 |
| Publisher Place | Brazil |
| Access Restriction | Open |
| Subject Keyword | Antigens, Cd55 Therapeutic Use Myasthenia Gravis, Autoimmune, Experimental Drug Therapy Neuroprotective Agents Animals Administration & Dosage Blood Disease Models, Animal Electrophoresis, Polyacrylamide Gel Enzyme-linked Immunosorbent Assay Injections, Intravenous Pathology Pharmacology Protein Renaturation Drug Effects Protein Structure, Tertiary Rats, Inbred Lew Recombinant Proteins Metabolism Solubility Discipline Genetics Discipline Molecular Biology Discipline Bioinformatics |
| Content Type | Text |
| Resource Type | Article |
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