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均匀设计与最小二乘支持向量机在转染优化中的集成应用。

Integrated application of uniform design and least-squares support vector machines to transfection optimization.

作者信息

Pan Jin-Shui, Hong Mei-Zhu, Zhou Qi-Feng, Cai Jia-Yan, Wang Hua-Zhen, Luo Lin-Kai, Yang De-Qiang, Dong Jing, Shi Hua-Xiu, Ren Jian-Lin

机构信息

Division of Gastroenterology, Zhongshan Hospital, Gastroenterology Institute of Xiamen University, Gastroenterology Center of Xiamen, Fujian Province, PR China.

出版信息

BMC Biotechnol. 2009 May 31;9:52. doi: 10.1186/1472-6750-9-52.

Abstract

BACKGROUND

Transfection in mammalian cells based on liposome presents great challenge for biological professionals. To protect themselves from exogenous insults, mammalian cells tend to manifest poor transfection efficiency. In order to gain high efficiency, we have to optimize several conditions of transfection, such as amount of liposome, amount of plasmid, and cell density at transfection. However, this process may be time-consuming and energy-consuming. Fortunately, several mathematical methods, developed in the past decades, may facilitate the resolution of this issue. This study investigates the possibility of optimizing transfection efficiency by using a method referred to as least-squares support vector machine, which requires only a few experiments and maintains fairly high accuracy.

RESULTS

A protocol consists of 15 experiments was performed according to the principle of uniform design. In this protocol, amount of liposome, amount of plasmid, and the number of seeded cells 24 h before transfection were set as independent variables and transfection efficiency was set as dependent variable. A model was deduced from independent variables and their respective dependent variable. Another protocol made up by 10 experiments was performed to test the accuracy of the model. The model manifested a high accuracy. Compared to traditional method, the integrated application of uniform design and least-squares support vector machine greatly reduced the number of required experiments. What's more, higher transfection efficiency was achieved.

CONCLUSION

The integrated application of uniform design and least-squares support vector machine is a simple technique for obtaining high transfection efficiency. Using this novel method, the number of required experiments would be greatly cut down while higher efficiency would be gained. Least-squares support vector machine may be applicable to many other problems that need to be optimized.

摘要

背景

基于脂质体的哺乳动物细胞转染对生物学专业人员来说是一项巨大挑战。为了抵御外源损伤,哺乳动物细胞往往表现出较差的转染效率。为了获得高效转染,我们必须优化转染的几个条件,如脂质体用量、质粒用量以及转染时的细胞密度。然而,这个过程可能既耗时又耗能。幸运的是,在过去几十年中发展起来的几种数学方法可能有助于解决这个问题。本研究探讨了使用一种称为最小二乘支持向量机的方法来优化转染效率的可能性,该方法只需要进行少量实验且能保持相当高的准确性。

结果

根据均匀设计原理进行了一个由15个实验组成的方案。在这个方案中,脂质体用量、质粒用量以及转染前24小时接种的细胞数量被设为自变量,转染效率被设为因变量。从自变量及其各自的因变量推导出一个模型。进行了另一个由10个实验组成的方案来测试该模型的准确性。该模型表现出很高的准确性。与传统方法相比,均匀设计和最小二乘支持向量机的综合应用大大减少了所需实验的数量。此外,还实现了更高的转染效率。

结论

均匀设计和最小二乘支持向量机 的综合应用是一种获得高转染效率的简单技术。使用这种新方法,所需实验的数量将大大减少,同时能获得更高的效率。最小二乘支持向量机可能适用于许多其他需要优化的问题。

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