Leemasawatdigul Krisada, Gappa-Fahlenkamp Heather
School of Chemical Engineering, Oklahoma State University, 423 Engineering North, Stillwater, OK 74078, USA.
Biologicals. 2011 Jan;39(1):29-32. doi: 10.1016/j.biologicals.2010.09.003. Epub 2010 Oct 20.
Monocyte chemoattractant protein-1 (MCP-1) is commercially available in a form of recombinant protein. This makes it more convenient to study the functions of MCP-1 and its involvement in many cell functions. However, when using MCP-1 in experimental studies, if the analysis is not performed immediately, the stability of recombinant MCP-1 may become an issue. In this study, the stability of recombinant MCP-1 at different concentrations and storage conditions was investigated. Results show that no significant loss of MCP-1 is observed when MCP-1 solutions were stored at non-freezing condition (4 °C) for seven days. However, for storage at freezing conditions (-20 °C or -81 °C), it appears that the first freeze-thaw cycle may contribute to some loss of MCP-1 in the solutions, and such loss may be concentration and time dependent. The effect of multiple freeze-thaw cycles for storage at freezing conditions was also examined. Data reveal that the second freeze-thaw cycle causes approximately 50% loss of MCP-1 in the solutions. This finding confirms that multiple freeze-thaw cycles should be avoided. The findings of this study provide an outline of how storage can affect the stability of recombinant proteins and should be taken into account during the evaluation of the concentration of recombinant proteins.
单核细胞趋化蛋白-1(MCP-1)有重组蛋白形式的市售产品。这使得研究MCP-1的功能及其在多种细胞功能中的作用更加便捷。然而,在实验研究中使用MCP-1时,如果不立即进行分析,重组MCP-1的稳定性可能会成为一个问题。在本研究中,对重组MCP-1在不同浓度和储存条件下的稳定性进行了研究。结果表明,当MCP-1溶液在非冷冻条件(4℃)下储存7天时,未观察到MCP-1有明显损失。然而,对于在冷冻条件(-20℃或-81℃)下储存,第一次冻融循环似乎可能导致溶液中MCP-1有所损失,且这种损失可能与浓度和时间有关。还研究了在冷冻条件下多次冻融循环的影响。数据显示,第二次冻融循环会导致溶液中约50%的MCP-1损失。这一发现证实应避免多次冻融循环。本研究结果概述了储存如何影响重组蛋白的稳定性,在评估重组蛋白浓度时应予以考虑。