Direct Comparison of Electric Field-Mediated and PEG-Mediated Cell Fusion for the Generation of Antibody Producing Hybridomas

Abstract

As an alternative to polyethylene glycol (PEG), electric field pulses offer, in theory, fusion conditions whose parameters are better controllable. In 1985 (1) we reported on the successful generation of hybridoma clones by means of electrofusion performed in a batch-type manner similar to that usually employed with PEG, and applicable to any type of antigens. Here we summarize the results of a series of fusions performed since then in which both electric field and PEG induced fusion were directly compared. Different types of antigens were used. Electrofusion resulted in a 3.8 to 33.0 times higher yield of hybridomas per unit number of spleen cells. Moreover, hybridomas grew more vigorously after fusion and, therefore, were earlier visible. Other parameters examined revealed no differences between hybridomas generated by either method.

Toxin–antigen conjugates as selection tools for antibody producing cells

“…But most of these techniques were not applicable as routine method or were just quantitative. Electrofusion generally increases the number of hybridoma clones after fusion up to more than 10 times (Karsten et al, 1988;Stoicheva and Hui, 1994). It is therefore applied in several laboratories but does increase the fusion rates not only of antigen-specific B lymphocytes but also of all cells.…”

Production of Monoclonal Antibodies by the Use of pH-Dependent Vesicular Stomatitis Virus-Mediated Cell Fusion

“…With regard to the current technology to fuse antibody-producing B cells with myeloma partner cells, the most popular technique is polyethylene glycol (PEG)-mediated cell fusion. PEG-fusion gives good yields of hybridomas if carefully controlled by following a detailed procedure, but the uncertainty with respect to the outcome of individual experiments still remains because the success of fusion depends, to some extent, on minute details which are difficult to standardize (1,4). Successful improvements of the fusion technique with electrofusion (2, 3, 4) and immunochemical methods (5, 6) have been reported, but none of these techniques have been widely adopted (1).…”

Droplet microfluidic platform for cell electrofusion

“…After counting on average 500 droplets per experiment (n = 5), of which 15% (75 droplets) contained two cells and qualified for possible electrofusion, 19 successful electrofusion events were found resulting in 5 % electrofusion efficiency. The established efficiency is comparable to the efficiency of both commercially available methods [53][54][55] as well as on-chip methods 52,[56][57][58] . To compete with existing methods, the electrofusion efficiency must be further improved, and there are several realistic opportunities for this.…”

“…The interaction of the induced dipole with a non-uniform electric field generates a DEP force (F DEP ), which is proportional to the cell volume (R 3 ), the field gradient E 0 and the real part of the Clausius-Mossotti (CM) factor, K(ω). F DEP is given by [53][54][55] :…”

“…by applying an AC electric field to provide a DEP force to improve cell alignment and cell-cell contact 55,93 ). In addition, a 3D configuration of the poration electrodes can be used to generate a homogeneous electric field along pairing axis of the cells, giving higher fusion yield.…”