柑桔速衰病毒株系特异性单克隆抗体的制备及其免疫学特性

PRODUCTION AND IMMUNOLOGICAL FEATURE OF SPECIFIC MONOCLONAL ANTIBODIES TO CITRUS TRISTEZA VIRUS STRAINS

以感染枸橼的柑桔速衰病毒(Citrus tristeza virus,CTV)的部分纯化制备物和浓缩的感染组织粗浸出液免疫BALB/C小白鼠后获得的脾脏细胞,与鼠骨髓瘤细胞株P3-X63-Ag8.653融合制备了杂交瘤细胞.免疫小鼠的CTV株系为T514,杂交瘤细胞以有限稀释法获得纯克隆,并以健康枸橼和CTV不同株系感染枸橼的组织浸出液筛选各克隆,并确认各克隆与CTV各株系的免疫反应性.经以碱性磷酸酶标记的山羊抗鼠IgG进行酶联免疫吸附试验(ELISA),对各杂交瘤克隆的株系特殊性分析证明,从这些杂交瘤细胞中获得了能产生CTV株系特异性单克隆抗体的杂交瘤细胞株.

Hybridomas are preparcd by the fusion of mouse myeloma cells, cell line P3-X63-Ag8. 653, and spleen cells from BALB/C mice immunized with partially clarified and concentration sap from Citrus tristeza virus （CTV） infected citron. A CTV strain, T514, is used for the preparation of inject antigcn. Hybrids are cloned by limiting dilution and screened for reactivity to sap from CTV infected and healthy citron. The analysis of clones for strain specificity is undertaken using a solid phase ELISA with alkaline phosphatase conjugated affinity purified goat anti-mouse IgG. The results of these tests indicate that strain specific or strain differentiating monoelonal antibodies might be obtained from these hybridomas.