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诊断血清学中使用的冻干辣根过氧化物酶结合单克隆抗体的稳定性

Stability of freeze dried horseradish peroxidase conjugated monoclonal antibodies used in diagnostic serology.

作者信息

Nielsen K

机构信息

Agriculture and Agri-Food Canada, Animal Diseases Research Institute, Nepean, Ontario, Canada.

出版信息

J Immunoassay. 1995 May;16(2):183-97. doi: 10.1080/15321819508013557.

Abstract

Two murine monoclonal antibodies, an IgG1 isotype specific for the heavy chain of bovine IgG1, the second an IgG3 isotype and specific for an epitope of the O-polysaccharide of Brucella abortus were conjugated with horseradish peroxidase. The conjugated antibodies were freeze dried in the presence of a number of additives to preserve activity and tested for stability over an 18 month period. Addition of 0.3M trehalose or 0.8% lactalbumin and 3.2% sucrose resulted in the lowest loss of activity if the conjugated antibodies were freeze dried in glass vials. Freeze drying in polypropylene vials resulted in a more rapid rate of deterioration with some additives or an accelerated rate of decline after an initial plateau of lesser loss of activity. The use of polypropylene vials and lactalbumin and sucrose were included in this study because of their low cost compared to glass vials and trehalose. While each antibody behaved differently, addition of trehalose or lactalbumin and sucrose to the conjugated antibodies aided in the preservation of enzyme activity. Both additives provide a suitable method for long term storage of freeze dried or freeze dried and reconstituted monoclonal antibody-enzyme conjugates.

摘要

两种鼠单克隆抗体,一种是针对牛IgG1重链的IgG1同型抗体,另一种是针对流产布鲁氏菌O-多糖表位的IgG3同型抗体,它们与辣根过氧化物酶偶联。将偶联抗体在多种添加剂存在下进行冷冻干燥以保持活性,并在18个月期间测试其稳定性。如果将偶联抗体在玻璃小瓶中冷冻干燥,添加0.3M海藻糖或0.8%乳白蛋白和3.2%蔗糖会导致活性损失最低。在聚丙烯小瓶中冷冻干燥,使用某些添加剂会导致降解速度更快,或者在活性损失较小的初始平稳期后下降速度加快。本研究中使用聚丙烯小瓶、乳白蛋白和蔗糖是因为与玻璃小瓶和海藻糖相比它们成本较低。虽然每种抗体的表现不同,但向偶联抗体中添加海藻糖或乳白蛋白和蔗糖有助于保持酶活性。这两种添加剂都为冷冻干燥或冷冻干燥并复溶的单克隆抗体-酶偶联物的长期储存提供了合适的方法。

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