Verheesen P, ten Haaft M R, Lindner N, Verrips C T, de Haard J J W
Department of Molecular and Cellular Biology, University of Utrecht, Utrecht 3500 TB, The Netherlands.
Biochim Biophys Acta. 2003 Dec 5;1624(1-3):21-8. doi: 10.1016/j.bbagen.2003.09.006.
We explored the possibility to apply single-domain antibodies from Camelidae for immunoaffinity purification of the ice structuring protein (ISP) from Lolium perenne, which modifies ice crystal growth and therefore has potential application in medicine, biotechnology, agriculture and (frozen) foods. Using phage display together with an appropriate selection method, a group of candidate fragments was isolated from a llama-derived immune library. Affinity chromatography using a purposely selected antibody coupled to a matrix yielded a completely pure and functional ISP. Due to the extreme refolding capabilities and physical stability of single-domain antibodies, the affinity matrix could be regenerated more than 2000 times without loss of capacity, while the fragment's monomeric nature permitted an efficient elution of antigen. The results of this study show that highly pure proteins can be recovered from biological material in a single-step process.
我们探索了应用骆驼科动物的单域抗体从多年生黑麦草中免疫亲和纯化冰结构蛋白（ISP）的可能性，该蛋白可改变冰晶生长，因此在医学、生物技术、农业和（冷冻）食品领域具有潜在应用价值。通过噬菌体展示结合适当的筛选方法，从羊驼来源的免疫文库中分离出一组候选片段。使用与基质偶联的特意选择的抗体进行亲和层析，得到了完全纯的且具有功能的ISP。由于单域抗体具有极强的重折叠能力和物理稳定性，亲和基质可重复使用2000多次而不损失容量，同时片段的单体性质允许高效洗脱抗原。本研究结果表明，可通过单步过程从生物材料中回收高纯度蛋白质。
