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Biotechnology and Applied Biochemistry (2002) 35, (171–180) (Printed in Great Britain)

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Effect of medium properties and additives on antibody stability and accumulation in suspended plant cell cultures

Bonny M.-Y. Tsoi and Pauline M. Doran¹

Department of Biotechnology, University of New South Wales, Sydney, NSW 2052, Australia

Key words: gibberellic acid, haemin, manganese, osmolality, tobacco.

Abbreviations used: ER, endoplasmic reticulum; MS medium, Murashige and Skoog medium; NAA, naphthaleneacetic acid; DMEM, Dulbecco's minimal essential medium; BiP, immunoglobulin heavy-chain binding protein.

¹To whom correspondence should be addressed (e-mail p.doran@unsw.edu.au).

Factors affecting antibody accumulation and stability were investigated in transgenic plant cell cultures. Whereas IgG₁ antibody was stably maintained in media used for animal cell culture, there was a rapid loss over a period of 1–2 h of antibody added to sterile plant culture media. Antibody stability in Gamborg's B5 medium was improved in the absence of Mn. Tobacco suspensions producing IgG₁ antibody were used to test various medium-based strategies for improving antibody accumulation in plant culture. Even though growth was suppressed, antibody levels in the biomass and medium were increased in media containing mannitol at osmolalities up to 450 mOsm · (kg of water)^-1. Adding gibberellic acid and haemin to the cultures was also beneficial, but the effects were not as great as those obtained under hyperosmolar conditions. Moderate increases in antibody accumulation were found by culturing plant cells in B5 medium without Mn.

Received 13 December 2001/21 February 2002; accepted 12 March 2002

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