Production of antibodies in transgenic plants.
Complementary DNAs derived from a mouse hybridoma messenger RNA were used to transform tobacco leaf segments followed by regeneration of mature plants. Plants expressing single gamma or kappa immunoglobulin chains were crossed to yield progeny in which both chains were expressed simultaneously. A functional antibody accumulated to 1.3% of total leaf protein in plants expressing full-length cDNAs containing leader sequences. Specific binding of the antigen recognized by these antibodies was similar to the hybridoma-derived antibody. Transformants having gamma- or kappa-chain cDNAs without leader sequences gave poor expression of the proteins. The increased abundance of both gamma- and kappa-chains in transformants expressing assembled gamma-kappa complexes was not reflected in increased mRNA levels. The results demonstrate that production of immunoglobulins and assembly of functional antibodies occurs very efficiently in tobacco. Assembly of subunits by sexual cross might be a generally applicable method for expression of heterologous multimers in plants.
Hiatt
A
A
Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Cafferkey
R
R
Bowdish
K
K
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D003433:Crosses, Genetic; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D005803:Genes, Immunoglobulin; D006825:Hybridomas / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007142:Immunoglobulin gamma-Chains / Q000235:genetics* / Q000302:isolation & purification; D007145:Immunoglobulin kappa-Chains / Q000235:genetics* / Q000302:isolation & purification; D010944:Plants / Q000235:genetics*; D010947:Plants, Toxic; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification; D014026:Nicotiana / Q000235:genetics; D014158:Transcription, Genetic; D014170:Transformation, Genetic
D007143:Immunoglobulin Heavy Chains; D007142:Immunoglobulin gamma-Chains; D007145:Immunoglobulin kappa-Chains; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004247:DNA