Hybridoma production by means of electrofusion — an experimental report

Abstract

For several years we have been using, besides the PEG technique, the method of electrofusion for hybridoma production. In our opinion, the latter method is clearly superior to the PEG technique in regard to hybridoma yield . A disadvantage of both methods is the randomness of pairing the fusion partners . To overcome this, use of the avidin-biotin system was proposed [1], and has also been tried by us. We can confirm the positive results in ref. 1. In addition, we have attempted, for the production of monoclonal antibodies against low-molar-mass compounds, to combine the advantages of electrofusion with an antigen-directed fusion, which is simpler relative to avidin-biotin coupling . Because of the great significance of monoclonal antibodies in biology and medicine their production has become very rapidly commercialized . However, a technologically optimized production does not yet exist . As is demonstrated in Fig . 1, the production of monoclonal antibodies is a complicated process of different steps. Animals must be immunized ; spleen cells removed and fused with myeloma cells ; hybridoma cells must be cloned ; and so on. Each step is a problem in itself . An essential step is cell fusion . The conventional technique is polyethylene glycol (PEG) mediated fusion, but the yield of fusion is low . Therefore, the discovery of electrofusion [2-4] was very interesting, because the yield of hybrid cells is distinctly better. These facts were discovered about 6 years ago . However, the number of hybrid cells is not sufficient to estimate the success of the fusion . Hybrid cells must be stably clonable, then they are qualified as viable clones [5-7] ; if they produce antibodies of high affinity to the antigen used, they are then called positive clones . 1 7 3