McCormick C A, Toll M O, Marshall W H
Faculty of Engineering, Memorial University of Newfoundland, St John's, Canada.
J Chem Technol Biotechnol. 1992;54(2):159-69. doi: 10.1002/jctb.280540210.
An important recent development in the field of biotechnology has been the use of high electric fields to render cell membranes temporarily permeable (electroporation). Cells in this state are receptive to gene transfer or can be induced to fuse with each other (electrofusion) to form hybrid cells containing the combined characteristics of the parent cells. A major reason for fusing cells is to form hybridoma cells which secrete monoclonal antibodies. A problem for research workers has been the high price of some of the electrofusion equipment. This problem has been addressed by designing a device that is inexpensive (less than $800 Canadian) and can be assembled by an electronics technician. The system uses a Radio Shack CoCo III microcomputer which is programmed in BASIC and controls the electroporation voltage pulse amplitude (25-500 V) and duration (2-275 microseconds) to an electrofusion chamber; this yields electric field strengths of 0.25-5 kV cm-1 for a 0.1 cm electrode spacing. The system is capable of delivering pulse currents up to 3 A. This paper provides technical details on how to construct an instrument that has greater flexibility at less cost than comparable commercially available instruments.
生物技术领域最近的一项重要进展是利用高电场使细胞膜暂时通透（电穿孔）。处于这种状态的细胞易于接受基因转移，或者可被诱导相互融合（电融合）以形成包含亲代细胞综合特征的杂交细胞。融合细胞的一个主要原因是形成分泌单克隆抗体的杂交瘤细胞。研究人员面临的一个问题是一些电融合设备价格高昂。通过设计一种价格低廉（低于800加元）且可由电子技术人员组装的装置，解决了这一问题。该系统使用一台Radio Shack CoCo III微型计算机，用BASIC语言编程，控制施加到电融合室的电穿孔电压脉冲幅度（25 - 500伏）和持续时间（2 - 275微秒）；对于0.1厘米的电极间距，这可产生0.25 - 5千伏/厘米的电场强度。该系统能够输送高达3安的脉冲电流。本文提供了有关如何构建一种仪器的技术细节，该仪器比同类市售仪器具有更大的灵活性且成本更低。
