We use cookies to improve your experience. By continuing to browse this site, you accept our cookie policy.×
Skip main navigation
Aging Health
Bioelectronics in Medicine
Biomarkers in Medicine
Breast Cancer Management
CNS Oncology
Colorectal Cancer
Concussion
Epigenomics
Future Cardiology
Future Medicine AI
Future Microbiology
Future Neurology
Future Oncology
Future Rare Diseases
Future Virology
Hepatic Oncology
HIV Therapy
Immunotherapy
International Journal of Endocrine Oncology
International Journal of Hematologic Oncology
Journal of 3D Printing in Medicine
Lung Cancer Management
Melanoma Management
Nanomedicine
Neurodegenerative Disease Management
Pain Management
Pediatric Health
Personalized Medicine
Pharmacogenomics
Regenerative Medicine

Hybridoma technologies for antibody production

    &
    Kanta Tsumoto

    Division of Chemistry for Materials, Graduate School of Engineering, Mie University, 1577 Kurima-Machiya-cho, Tsu, Mie 514-8507, Japan

    Published Online:https://doi.org/10.2217/imt.11.4

    Hybridoma technology features effective usage of innate functions of both immune cells and cancers, allowing production of hybridoma cells, which continuously generate monoclonal antibodies specific to antigens of interest. For standard generation of hybridoma cells, B lymphocytes must be somatically fused with myeloma cells using various technologies. However, the methods generally do not necessarily result in selective fusion of target B lymphocytes with myeloma cells. To overcome this problem, we have developed a new hybridoma technology that involves preselection of B lymphocytes with target antigens based on immunoglobulin receptors and selective fusion of B cell–myeloma cell complexes with electrical pulses. The advanced methodology, termed B-cell targeting, multitargeting and stereospecific targeting, may be applicable to simultaneous production of monoclonal antibodies, selective production of stereospecific monoclonal antibodies, and also to efficient generation of human monoclonal antibodies for clinical purposes.

    Papers of special note have been highlighted as: ▪ of interest ▪▪ of considerable interest

    Bibliography

    • K öhler G, Milstein C: Continuous cultures of fused cells secreting antibody of predefined specificity. Nature256(5517),495–497 (1975).▪▪ Original hybridoma technology for the production of monoclonal antibodies by fusing B lymphocytes with cancerous myeloma cells.
    • Benjamini E, Coico R, Sunshine G: Immunology, a Short Course (Fourth Edition). Wiley–Blackwell, NY, USA (2000).
    • de StGroth SF, Scheidegger D: Production of monoclonal antibodies: strategy and tactics. J. Immunol. Methods35(1–2),1–21 (1980).
    • Zimmermann U: Electric field-mediated fusion and related electrical phenomena. Biochim. Biophys. Acta694(3),227–277 (1982).▪ Demonstrated improved hybridoma technology by use of pearl-chain formation. The technique brought about relatively high fusion efficiency.
    • Ohkohchi N, Itagaki H, Doi H, Taguchi Y, Satomi S, Satoh S: New technique for producing hybridoma by using laser radiation. Lasers Surg. Med.27(3),262–268 (2000).▪ Another improved hybridoma technology using pulse laser, which enabled manually controlled cell fusion under a microscope.
    • Lo MMS, Tsong TY, Conrad MK, Strittmatter SM, Hester LD, Snyder SH: Monoclonal antibody production by receptor-mediated electrically induced cell fusion. Nature310(5980),792–794 (1984).▪▪ Original report for the next generation of hybridoma technology based on antigen-based preselection of B lymphocytes and electrical pulses.
    • Tsong TY, Tomita M, Lo MMS: Pre-selection of B-lymphocytes by antigen for fusion to myeloma cells by pulsed electric field (PEF) method. In: Molecular Mechanisms of Membrane Fusion. Ohki S, Doyle D, Flanagan TD, Hui SW, Mayhew E (Eds). Plenum, NY, USA, 223–236 (1988).
    • Tomita M, Tsong TY: Selective production of hybridoma cells: antigenic-based pre-selection of B lymphocytes for electrofusion with myeloma cells. Biochim. Biophys. Acta1055(3),199–206 (1990).
    • Tsong TY, Tomita M: Selective B lymphocyte-myeloma cell fusion. Methods Enzymol.220,238–246 (1993).
    • 10  Tomita M, Sugi H, Ozawa K, Tsong TY, Yoshimura T: Targeting antigen-specific receptors on B lymphocytes to generate high yields of specific monoclonal antibodies directed against biologically active lower antigenic peptides within presenilin 1. J. Immunol. Methods251(1–2),31–43 (2001).
    • 11  Tomita M, Fukuda T, Ozu A, Kimura K, Tsong TY, Yoshimura T: Antigen-based immunofluorescence analysis of B-cell targeting: advanced technology for the generation of novel monoclonal antibodies with high efficiency and selectivity. Hybridoma (Larchmt) 25(5),283–292 (2006).▪ Three critical steps of B-cell targeting were first corroborated on the basis of immunofluorescence analysis.
    • 12  Tomita M: Next generation of monoclonal antibodies. BIOforum Europe12(4),33–34 (2008).
    • 13  Tomita M: A novel method for generation of monoclonal antibodies. In: Biochemistry and Histocytochemistry Research Developments (Expert Commentary). Fuchs S, Auer M (Eds). Nova Science Publishers, NY, USA, 335–336 (2010).
    • 14  Tomita M, Tsumoto K: Recent advances in antigen-based generation of monoclonal antibodies. Curr. Immunol. Rev.6(1),56–61 (2010).▪ Review of antigen-based generation of monoclonal antibodies.
    • 15  Chaiet L, Wolf FJ: The properties of streptavidin, a biotin-binding protein produced by streptomycetes. Arch. Biochem. Biophys.106,1–5 (1964).
    • 16  Green NM: Avidin. Adv. Protein Chem.29,85–133 (1975).
    • 17  Wojchowski DM, Sytkowski AJ: Hybridoma production by simplified avidin-mediated electrofusion. J. Immunol. Methods90(2),173–177 (1986).
    • 18  Hewish DR, Werkmeister JA: The use of an electroporation apparatus for the production of murine hybridomas. J. Immunol. Methods120(2),285–289 (1989).
    • 19  Werkmeister JA, Tebb TA, Kirkpatrick A, Shukla DD: The use of peptide-mediated electrofusion to select monoclonal antibodies directed against specific and homologous regions of the potyvirus coat protein. J. Immunol. Methods143(2),151–157 (1991).
    • 20  Woolley DW, Longsworth LG: Isolation of an antibiotin factor from egg white. J. Biol. Chem.142(1),285–290 (1942).
    • 21  Tomita M, Yokota T, Wada M et al.: Nanosensing measurement using novel monoclonal antibodies generated by B-cell targeting. FEBS J.275(Suppl. 1),373 (2008).
    • 22  Boss BD: An improved in vitro immunization procedure for the production of monoclonal antibodies. Methods Enzymol.121,27–33 (1986).
    • 23  Tomita M, Taniguchi T, Tanaka J et al.: Generation of monoclonal antibodies directed against peptide sequences by a B cell targeting technique. Hum. Antibodies12(1–2),37–39 (2003).
    • 24  Tomita M, Sugao M, Taniguchi T et al.: Antigen-based B cell targeting technique to generate high yields of monoclonal antibodies. Hum. Antibodies13(1–2),42 (2004).
    • 25  Tomita M, Taniguchi T, Sawada H, Nishino N, Yoshimura T: Rapid generation of monoclonal antibodies against the epitope sequences of human insulin using an in vitro immunization method. Peptide Sci.469–472 (2004).
    • 26  Tomita M, Asaoka Y, Kato Y, Ogata S, Tsong TY, Yoshimura T: A new technology for the generation of novel monoclonal antibodies based on short-term immunization. Peptide Sci.105–108 (2006).
    • 27  Tomita M, Asaoka Y, Fukuda T et al.: Rapid and selective production of hybridoma cells secreting novel monoclonal antibodies based on short-term immunization. Hum. Antibodies15(1–2),33–35 (2006).
    • 28  Gavilondo JV, Larrick JW: Antibody engineering at the millennium. BioTechniques29(1),128–145 (2000).
    • 29  Konthur Z, Hust M, Dübel S: Perspectives for systematic in vitro antibody generation. Gene364,19–29 (2005).
    • 30  Wark KL, Hudson PJ: Latest technologies for the enhancement of antibody affinity. Adv. Drug Deliv. Rev.58(5–6),657–670 (2006).
    • 31  Bradbury AR, Marks JD: Antibodies from phage antibody libraries. J. Immunol. Methods290(1–2),29–49 (2004).
    • 32  Border ET, Midelfort KS, Wittrup KD: Directed evolution of antibody fragments with monovalent femtomolar antigen-binding affinity. Proc. Natl Acad. Sci. USA97(20),10701–10705 (2000).
    • 33  Feldhaus MJ, Siegel RW: Yeast display of antibody fragments: a discovery and characterization platform. J. Immunol. Methods290(1–2),69–80 (2004).
    • 34  Lipovsek D, Plückthun A: In-vitro protein evolution by ribosome display and mRNA display. J. Immunol. Methods290(1–2),51–67 (2004).
    • 35  Løvborg U: Monoclonal Antibodies: Production and Maintenance. William Heinemann Medical Books, London, UK (1982).
    • 36  Tomita M, Furuta K, Sano Y, Tsumoto K: Multiple generation of novel monoclonal antibodies based on B-cell targeting. FEBS J.276(Suppl. 1),293 (2009).
    • 37  TOMITA M, MATSUBA T: JP4599527 (2010).
    • 38  Tomita M, Hirano M, Watanabe E et al.: A new technology for intact antigen-based and receptor-mediated generation of novel monoclonal antibodies. Hum. Antibodies16(1–2),40–41 (2007).
    • 39  Tomita M, Tsumoto K: New hybridoma technology based on antigen-specific immunoglobulin receptors. FEBS J.277(Suppl. 1),50 (2010).
    • 40  Lonberg N, Taylor LD, Harding FA et al.: Antigen-specific human antibodies from mice comprising four distinct genetic modifications. Nature368(6474),856–859 (1994).
    • 41  Green LL, Hardy MC, Maynard-Currie CE et al.: Antigen-specific human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs. Nat. Genet.7(1),13–21 (1994).
    • 42  Jakobovits A: Production of fully human antibodies by transgenic mice. Curr. Opin. Biotechnol.6(5),561–566 (1995).
    • 43  Matsumoto N, Matsue T, Uchida I: Paired cell alignment using jagged microarray electrode. Bioelectrochem. Bioenerg.34(2),199–202 (1994).
    • 44  Ho CT, Lin RZ, Chang WY, Chang HY, Liu CH: Rapid heterogeneous liver-cell on-chip patterning via the enhanced field-induced dielectrophoresis trap. Lab. Chip6(6),724–734 (2006).
    • 45  Heida T, Rutten WLC, Marani E: Dielectrophoretic trapping of dissociated fetal cortical rat neurons. IEEE Trans. Biomed. Eng.48(8),921–930 (2001).
    • 46  Gray DS, Tan JL, Voldman J, Chen CS: Dielectrophoretic registration of living cells to a microelectrode array. Biosens. Bioelectron.19(12),1765–1774 (2004).
    • 47  Yu Z, Xiang G, Pan L et al.: Negative dielectrophoretic force assisted construction of ordered neuronal networks on cell positioning bioelectronic chips. Biomed. Microdevices6(4),311–324 (2004).
    • 48  Suzuki M, Yasukawa T, Shiku H, Matsue T: Negative dielectrophoretic patterning with different cell types. Biosens. Bioelectron.24(4),1043–1047 (2008).
    • 49  Jat PS, Noble MD, Ataliotis P et al.: Direct derivation of conditionally immortal cell lines from an H-2Kb-tsA58 transgenic mouse. Proc. Natl Acad. Sci. USA88(12),5096–5100 (1991).
    • 50  Pasqualini R, Arap W: Hybridoma-free generation of monoclonal antibodies. Proc. Natl Acad. Sci. USA101(1),257–259 (2004).
    • 51  Matsuba T, Mitoma Y, Tanaka C, Inoue M: Preparation of super-high affinity rabbit monoclonal antibodies against estradiol. Application to highly sensitive estradiol measurement. Tosoh Res. Technol. Rev.52,3–9 (2008).
    • 52  MATSUBA T: JP2009240300 (2009).
    • 53  MASUBUCHI H, HASHIMOTO S: JP1998262657 (1998).
    • 54  Zhou X, Hu W, Qin X: The role of complement in the mechanism of action of rituximab for B-cell lymphoma: implications for therapy. Oncologist13(9),954–966 (2008).
    • 55  Kubota T, Niwa R, Satoh M, Akinaga S, Shitara K, Hanai N: Engineered therapeutic antibodies with improved effector functions. Cancer Sci.100(9),1566–1572 (2009).