Expression and characterization of a recombinant single-domain monoclonal antibody against MUC1 mucin in tobacco plants.
A promising alternative to conventional antibodies is the single-domain antibody fragment of the Camelidae (V(HH)), which (because of features such as small length, high expression, solubility, and stability) is preferred to other antibody derivatives. In this report, a recombinant single-domain antibody (V(HH)) against MUC1 mucin in the tobacco plant, which may be considered as a suitable and economical alternative expression system, was produced. This antibody was expressed under the control of a strong constitutive promoter, CaMV35S, and NOS terminator. A plant high-expression sequence (Kozak sequence) was linked at the 5' end for overexpression of the V(HH) gene. The constructed cassette (pBIV(HH)) was transferred to agrobacterium, and the VHH gene was inserted into the plant genome by agrobacterium-mediated transformation. Transgenic lines were selected on kanamycin (100 mg/L) and maintained in soil, and subsequent generations were obtained. The presence and expression of the transgene was confirmed in the transformants by polymerase chain reaction (PCR), enzyme-linked immunosorbent assay (ELISA), and Western blot. Tobacco transgenic lines leave expressed V(HH) at levels varying from 1.12% to 1.63% of the total soluble protein. This report examines the transformation and expression of recombinant single-domain antibody (V(HH)) against antigen-associated tumor in tobacco plants.
Rajabi-Memari
H
H
Department of Plant Breeding, School of Agriculture, Tarbiat Modares University, Tehran, Iran.
Jalali-Javaran
M
M
Rasaee
M J
MJ
Rahbarizadeh
F
F
Forouzandeh-Moghadam
M
M
Esmaili
A
A
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics*; D006825:Hybridomas; D018396:Mucin-1 / Q000276:immunology*; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D018396:Mucin-1; D011994:Recombinant Proteins