Production of an anti-mouse MHC class II monoclonal antibody with biological activity in transgenic tobacco.
To produce a monoclonal antibody specific to a mouse major histocompatibility complex (MHC) class II protein, we synthesized the complementary DNAs for the heavy and light chains of a monoclonal antibody by PCR amplification. These cDNAs were then introduced separately into tobacco plant cells. After performing Northern blot analysis to confirm the expression of each of the chain genes in the transformed plants, we constructed transgenic plants expressing both the heavy and light chains by sexual crossing. The expression of the heavy and light chain genes in the sexually crossed plant was confirmed by Northern and Western blot analyses, respectively. Fluorocytometric analysis showed that the plant-derived antibodies, which we purified using a protein G affinity column, bound specifically to target cells that expressed the cognate MHC class II molecules on their cell surfaces. The results of this study demonstrate that a monoclonal antibody against mouse MHC class II proteins can be expressed in transgenic plants. They also show the specific binding activity of plant-derived antibodies to cognate antigens.
Hong
Shin-Young
SY
Division of Biological Sciences, Research Center for Bioactive Materials, Chonbuk National University, Jeonju 561-756, Republic of Korea.
Kim
Tae-Geum
TG
Kwon
Tae-Ho
TH
Jang
Yong-Suk
YS
Yang
Moon-Sik
MS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D015152:Blotting, Northern; D015153:Blotting, Western; D018076:DNA, Complementary / Q000235:genetics; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000032:analysis / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000032:analysis / Q000235:genetics; D051379:Mice; D030821:Plants, Genetically Modified / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D014026:Nicotiana / Q000235:genetics*
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D000949:Histocompatibility Antigens Class II; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D012333:RNA, Messenger