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Recombinant expression of nanobodies and nanobody-derived immunoreagents.
Protein Expression and Purification ( IF 1.4 ) Pub Date : 2020-04-11 , DOI: 10.1016/j.pep.2020.105645
Ario de Marco 1
Affiliation  

Antibody fragments for which the sequence is available are suitable for straightforward engineering and expression in both eukaryotic and prokaryotic systems. When produced as fusions with convenient tags, they become reagents which pair their selective binding capacity to an orthogonal function. Several kinds of immunoreagents composed by nanobodies and either large proteins or short sequences have been designed for providing inexpensive ready-to-use biological tools. The possibility to choose among alternative expression strategies is critical because the fusion moieties might require specific conditions for correct folding or post-translational modifications. In the case of nanobody production, the trend is towards simpler but reliable (bacterial) methods that can substitute for more cumbersome processes requiring the use of eukaryotic systems. The use of these will not disappear, but will be restricted to those cases in which the final immunoconstructs must have features that cannot be obtained in prokaryotic cells. At the same time, bacterial expression has evolved from the conventional procedure which considered exclusively the nanobody and nanobody-fusion accumulation in the periplasm. Several reports show the advantage of cytoplasmic expression, surface-display and secretion for at least some applications. Finally, there is an increasing interest to use as a model the short nanobody sequence for the development of in silico methodologies aimed at optimizing the yields, stability and affinity of recombinant antibodies.

中文翻译:

纳米抗体和纳米抗体衍生的免疫试剂的重组表达。

可获得该序列的抗体片段适合在真核和原核系统中直接进行工程设计和表达。当与方便的标签融合生产时,它们成为将其选择性结合能力与正交功能配对的试剂。由纳米抗体和大蛋白或短序列组成的几种免疫试剂已被设计用于提供廉价的即用型生物学工具。在替代表达策略中进行选择的可能性至关重要,因为融合部分可能需要特定条件才能正确折叠或翻译后修饰。在纳米抗体生产的情况下,趋势是朝着更简单但可靠(细菌)的方法发展,该方法可以替代需要使用真核系统的繁琐过程。这些的使用不会消失,但将限于最终的免疫结构必须具有原核细胞无法获得的特征的情况。同时,细菌表达已经从常规方法演变而来,该常规方法仅考虑周质中的纳米抗体和纳米抗体融合体的积累。一些报告显示了至少在某些应用中细胞质表达,表面展示和分泌的优势。最后,人们越来越有兴趣使用短的纳米抗体序列作为模型来开发计算机模拟方法,以优化重组抗体的产量,稳定性和亲和力。但是将限于最终免疫结构必须具有原核细胞无法获得的特征的情况。同时,细菌表达已经从常规方法演变而来,该常规方法仅考虑周质中的纳米抗体和纳米抗体融合体的积累。一些报告显示了至少在某些应用中细胞质表达,表面展示和分泌的优势。最后,人们越来越有兴趣使用短的纳米抗体序列作为模型来开发计算机模拟方法,以优化重组抗体的产量,稳定性和亲和力。但是将限于最终免疫结构必须具有原核细胞无法获得的特征的情况。同时,细菌表达已经从常规方法演变而来,该常规方法仅考虑周质中的纳米抗体和纳米抗体融合体的积累。一些报告显示了至少在某些应用中细胞质表达,表面展示和分泌的优势。最后,人们越来越有兴趣使用短的纳米抗体序列作为模型来开发计算机模拟方法,以优化重组抗体的产量,稳定性和亲和力。一些报告显示了至少在某些应用中细胞质表达,表面展示和分泌的优势。最后,人们越来越有兴趣使用短的纳米抗体序列作为模型来开发计算机模拟方法,以优化重组抗体的产量,稳定性和亲和力。一些报告显示了至少在某些应用中细胞质表达,表面展示和分泌的优势。最后,人们越来越有兴趣使用短的纳米抗体序列作为模型来开发计算机模拟方法,以优化重组抗体的产量,稳定性和亲和力。
更新日期:2020-04-12
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