Preparation and Charaterization of Monoclonal Antibodies Against Recent Newcastle Disease Virus Isolate in Taiwan.
| Autor: | Gong Jia-yn, 龔加吟 |
|---|---|
| Rok vydání: | 2002 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 90 Fourteen clones of monoclonal antibody (mAb) production by hybridoma cell were prepared from 3 times fusion of the BALB/c mice spleen cell preimmunized with a 1999 local isolate (V157 strain) of genotype Ⅶ viscerotropic velogenic Newcastle disease virus (vvNDV) and mice myeloma cell. The production of mAbs against V157 strain in the hybridoma cell suspension or mice ascites were screened by enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody assay (IFA), then characterized by immuno dot blotting assay (ID), Western blot (WB) and neutralization test (NT). All clones of mAb were positive in ELISA and IFA. Only 1, 7, 3 and 2 clones of mAb showed positive at 4, 8, 16 and 512 dilution folds level, respectively. Ten (72％) clones of mAb recognized the linear epitope and the other 4 (28％) recognized the conformational dependent epitope on F-glycoprotein. At subtype level, 5, 3, 2 and 4 clones of mAb belonged to IgG1, IgG2a, IgG2b and IgG3, respectively. By competitive ELISA classification, 6, 5, 2 and 1 clones of mAb belong to A, B, C and D group, respectively. Which implys at least 4 antigenic epitopes existing on the F-glycoprotein of V157 strain. The conformation-dependent epitope was found only showing in group B and D. Although the production of NDV mAb is time consuming, the high specificity of mAb in binding with its special NDV antigen make the preparation of NDV mAb is very worthful studing the variation on the antigenic F-glycoprotein epitope in new isolate of vvNDV. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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