Improvement in the basic technology of electrofusion for generation of antibody-producing hybridomas

Ohnishi, Kouhei; Chiba, Joe; Goto, Yoji; Tokunaga, Tatsuya

doi:10.1016/0022-1759(87)90188-8

Cited by 58 publications

(11 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The work presented here illustrates that judicious application of the principles of electric field mediated cell fusion and standard immunization techniques are sufficient to obtain large numbers of antibody-secreting hybridomas. Moreover, our frequencies of specific hybridoma generation were comparable or superior to those reported using prefusion manipulations (15,21,22).…”

Section: Introductionsupporting

confidence: 81%

“…Thus analysis of field line distribution around dielectrophoretically aligned cells (30), study of membrane changes during the electrofusion process (31), new immunization techniques and the addition of novel fusion enhancing agents (15) and fusion under microgravity conditions (32) may permit additional gains in efficiency.…”

Section: Discussionmentioning

confidence: 99%

“…and NASA (NAG 82716) and the Ludwig Institute for Cancer Research to G.A.N. antigen bridging strategies (21,22) which have permitted dramatically increased numbers of antibody forming hybridomas, while Ohnishi et al (15) have obtained improved fusion yields by pretreating the fusion partners with proteases and by adding "facilitators" such as PEG, calmodulins, and glycerol monooleate.…”

Section: Introductionmentioning

confidence: 98%

See 2 more Smart Citations

Efficient Generation of Stable Antibody Forming Hybridoma Cells by Electrofusion

Schmitt¹,

Zimmermann²,

Neil³

1989

Hybridoma

View full text Add to dashboard Cite

A variety of modified electrofusion protocols designed to improve the efficiency of hybridoma production have recently appeared in the literature. We undertook to maximize the number of antibody secreting murine hybridomas by optimizing the temperature and fusion strength parameters of the conventional electrofusion technique. Anti-DNP secreting hybridomas were generated by fusing SP2/0 to immunized mouse splenic lymphocytes using an unmodified electrofusion protocol consisting of washing in a weakly conducting sorbitol fusion medium supplemented with bovine serum albumin, calcium and magnesium ions. This was followed by dielectrophoretic alignment and application of 3 short duration, high intensity field pulses in helical chambers. Optimal efficiencies of hybridomas were generated by the application of 2000 V/cm pulses at 25 degrees C (2.45 hybridomas x 10(-4) splenocytes) and as many as 63% of resulting hybridomas secreted anti-DNP monoclonal antibodies, the majority of which were IgG's. These data show that modification of the electrofusion protocol by pretreatment of fusion partners with proteolytic enzymes or the use of antigen bridging is not required for the successful and efficient production of specific monoclonal antibodies by electrofusion.

show abstract

Section: Introductionsupporting

confidence: 81%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

See 1 more Smart Citation

Efficient Generation of Stable Antibody Forming Hybridoma Cells by Electrofusion

Schmitt¹,

Zimmermann²,

Neil³

1989

Hybridoma

View full text Add to dashboard Cite

show abstract

“…Recently, several new methods of fusion have been reported, such as those using electrofusion (19)(20) or using non-serum medium (28). These methods are especially useful to produce HMA-secreting hybridomas.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, there was a total of 1-2x105 cells/well. 2) Electrofusion (19)(20): In some instances, lymphocytes were fused with parental cells by electrofusion.…”

Section: Fusion Proceduresmentioning

confidence: 99%

Characterization of an Established Mouse-Human Heterohybridoma and its Application for Production of (Mouse-Human)-Human Triple Hybridoma Secreting Human Immunoglobulin

Sugiyama¹,

Goto²,

Saito³

et al. 1991

Hybridoma

View full text Add to dashboard Cite

We report the construction of a mouse-human (M-H) heterohybridoma by fusion of the murine myeloma cell line NS-1 and human spleen cells from a 17 week old fetus. The nonsecreting, cloned hybridoma cell line II was resistant to 8-azaguanine (8-AG) and sensitive to hypoxanthine, aminopterin and thymidine (HAT) medium. It grew rapidly in 8-AG containing medium (doubling time 20 hrs.), but did not grow in HAT medium or in non-serum medium. It had a high fusion frequency with human lymphocytes from regional lymph nodes. Five human chromosomes were retained stably for over 6 months by this cell line II. Nine (mouse-human)-human ((M-H)-H) triple hybridomas secreting human IgG 1 or IgM were established by the fusion of this parental cell line II and human lymphocytes from regional lymph nodes. Immunoglobulin secretion was stable and has been maintained for over 8-10 months without recloning in these hybridomas. Secretion of immunoglobulin varies from 2.1-3.0 micrograms/10(6) cells/day, and these hybridomas contain from 3 to 16 human chromosomes, including No. 14. So, this M-H heterohybridoma II is an excellent useful parental cell line for the production of hybridomas secreting human immunoglobulin.

show abstract

Selection Strategies I: Monoclonal Antibodies

Moldenhauer¹

2007

Handbook of Therapeutic Antibodies

View full text Add to dashboard Cite

Improvement in the basic technology of electrofusion for generation of antibody-producing hybridomas

Cited by 58 publications

References 36 publications

Efficient Generation of Stable Antibody Forming Hybridoma Cells by Electrofusion

Efficient Generation of Stable Antibody Forming Hybridoma Cells by Electrofusion

Characterization of an Established Mouse-Human Heterohybridoma and its Application for Production of (Mouse-Human)-Human Triple Hybridoma Secreting Human Immunoglobulin

Selection Strategies I: Monoclonal Antibodies

Contact Info

Product

Resources

About