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猪IgE的分离及多克隆抗血清的制备。

Isolation of porcine IgE, and preparation of polyclonal antisera.

作者信息

Roe J M, Patel D, Morgan K L

机构信息

Department of Veterinary Medicine, University of Bristol, Langford, UK.

出版信息

Vet Immunol Immunopathol. 1993 Jul;37(2):83-97. doi: 10.1016/0165-2427(93)90057-b.

Abstract

High Titre, Ascaris specific, porcine reaginic antibody was induced by combining parasitic infestation and antigen challenge. The reagin was characterised as being heat labile at 56 degrees C, possessing long term skin sensitising activity, and as having physicochemical properties similar to IgE in other species. Porcine IgE was purified by conventional immunochemical techniques, and an epsilon chain isolate prepared using SDS-PAGE gel excision and electro elution. Antisera raised against the intact IgE molecule and its epsilon chain were rendered monospecific by immunoabsorption and characterised by reverse cutaneous anaphylaxis (RCA) testing, passive cutaneous anaphylaxis inhibition, immunohistology and Western blotting. These antisera are currently being used to study the regulation of IgE responses in the pig.

摘要

通过将寄生虫感染与抗原刺激相结合,诱导产生了高滴度的、针对蛔虫的猪反应素抗体。该反应素的特点是在56摄氏度时对热不稳定,具有长期皮肤致敏活性,并且其理化性质与其他物种的IgE相似。通过传统免疫化学技术纯化猪IgE,并使用SDS-PAGE凝胶切除和电洗脱制备ε链分离物。通过免疫吸附使针对完整IgE分子及其ε链产生的抗血清具有单特异性,并通过反向皮肤过敏反应(RCA)测试、被动皮肤过敏反应抑制、免疫组织学和蛋白质印迹进行表征。这些抗血清目前正用于研究猪体内IgE反应的调节。

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