In Vitro and In Vivo Immunization Techniques for the Production of Monoclonal Antibodies against the Aster Yellows Phytoplasma

以試管內及生體內免疫法研製翠菊黃化病菌質體之單元抗體

林長平(Chan-Pin Lin) ； 陳澤安(Tseh-An Chen)

《植物病理學會刊》 5卷1期 (1996/03) Pp. 28-32

https://doi.org/10.6649/PPB.199603_5(1).0003

摘要

本研究乃利用試管內及生體內免疫法進行翠菊黃化病菌質體單元抗體之研製，所採用之免疫抗原乃利用Percoll 梯度離心法純化所得，此種方法所製備之抗原比先前報告中利用由媒介昆蟲唾液腺所提取之抗原為佳，能大幅提升對翠菊黃化病菌質體具特異性抗體之融合瘤在所有具抗體產生能力之融合瘤族群中之比例，而且試管內免疫方式對此一比例之提升更為有利。在本一研究中共獲得十六株具特異性之融合瘤株，其所產生之單元抗體可利用ELISA法區分其他七種植物菌質髓體。

關鍵字

單元抗體 ； 試管內免疫法

並列摘要

In vitro and in vivo immunization techniques were applied for the production of monoclonal antibodies against the aster yellows (AY) agent, a phytoplasma. The immunogens purified from diseased lettuce using discontinuous Percoll density gradients, when compared with those directly extracted from the salivary glands of infective insect vectors applied in the previous study, enhanced the proportion of AY-specific antibody-producing hybridomas in the total antibody-secreting hybridoma populations in both in vitro and in vivo trials. The in vitro immunization, when compared with in vivo immunization, yielded an even higher proportion of AV-specific antibody-producing hybridomas. Sixteen stable hybridoma cell lines were produced, each secreting specific monoclonal antibodies against the AY agent . When using indirect biotinylated enzyme -linked immunosorbent assay (ELISA) , these monoclonal antibodies reacted specifically to the AY-infected plants and differentiated the AY agent from other phytoplasmas associated with seven other diseases.

並列關鍵字

monoclonal antibody ； in vitro immunization