Assembly of an antibody and its derived antibody fragment in Nicotiana and Arabidopsis.
The yield and assembly of an IgG1 antibody and its derived F(ab) fragment were compared in Nicotiana and Arabidopsis. The results obtained showed a lot of interclonal variability. For 45% of the primary transgenic calluses, antigen-binding entities represented less than 0.1% of the total soluble protein (TSP). Only two of the 103 analysed transformants contained more than 1% of antigen-binding protein, with 1.26% being the highest yield. Analogous amounts of complete antibody and F(ab) accumulated in primary callus tissue. Moreover, yields were in the same range for both species as far as primary callus tissue is concerned. However, the accumulation of the F(ab) fragment in leaf tissue of regenerated plants differed significantly between Nicotiana and Arabidopsis. The F(ab) fragment accumulated to only 0.044% of TSP in Nicotiana leaves but up to 1.3% in Arabidopsis leaves. Furthermore, both species showed differences in the assembly pattern of the complete antibody. Whereas Arabidopsis contained primarily fully assembled antibodies of 150 kDa, Nicotiana showed an abundance of fragments in the 50 kDa range.
De Neve
M
M
Laboratorium voor Genetica, Universiteit Gent, Belgium.
De Loose
M
M
Jacobs
A
A
Van Houdt
H
H
Kaluza
B
B
Weidle
U
U
Van Montagu
M
M
Depicker
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics*; D000917:Antibody Formation; D000937:Antigen-Antibody Reactions; D017360:Arabidopsis / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D007128:Immunoglobulin Fragments / Q000235:genetics*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G