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Plant Physiol, September 2000, Vol. 124, pp. 173-182
Effect of Climate Conditions and Plant Developmental Stage on the
Stability of Antibodies Expressed in Transgenic Tobacco
Lucas H.
Stevens,*
Geert M.
Stoopen,
Ingrid J.W.
Elbers,
Jos W.
Molthoff,
Hans A.C.
Bakker,
Arjen
Lommen,
Dirk
Bosch, and
Wilco
Jordi
Plant Research International, P.O. Box 16, NL-6700 AA Wageningen,
The Netherlands (L.H.S., G.M.S., J.W.M., H.A.C.B., D.B., W.J.); and
State Institute for Quality Control of Agricultural Products (RIKILT),
P.O. Box 230, NL-6700 AE Wageningen, The Netherlands (I.J.W.E.,
A.L.)
Plants are regarded as a promising system for the production of
heterologous proteins. However, little is known about the influence of
plant physiology and plant development on the yield and quality of the
heterologous proteins produced in plants. To investigate this, tobacco
(Nicotiana tabacum cv Samsun NN) was transformed with a
single construct that contained behind constitutive promotors the
light- and heavy-chain genes of a mouse antibody. The in planta
stability of the antibody was analyzed in transgenic plants that were
grown under high and low irradiation at 15°C and 25°C. High-light
conditions favored the production of biomass, of total soluble protein,
and of antibody. The plants grown at 25°C developed faster and
contained less antibody per amount of leaf tissue than the plants grown
at 15°C. Both endogenous protein and antibody content showed a strong
decline during leaf development. The heavy chains of the antibody
underwent in planta degradation via relatively stable fragments. In
vitro incubations of purified plantibody with leaf extracts of
wild-type tobacco indicated the involvement of acidic proteases. It is
interesting that the same antibody produced by mouse hybridoma cells
exhibited higher stability in this in vitro assay. This may be
explained by the assumption that the plant type of
N-glycosylation contributes less to the stability of the
antibody than the mouse-type of N-glycosylation. The
results of this study indicate that proteolytic degradation during
plant development can be an important factor affecting yield and
homogeneity of heterologous protein produced by transgenic plants.
*
Corresponding author; e-mail l.h.stevens{at}plant.wag-ur.nl;
fax 31-317-42-31-10.
© 2000 American Society of Plant Physiologists
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