The ErbB2 (Her2/neu) gene encodes a 185-kDa glycoprotein that is ligand-less receptor with a tyrosine kinase activity. It is overexpressed in human cancers such as breast, lung, kidney, bladder and ovary carcinomas, while it is low expressed in normal tissues. Structural studies demonstrated that oncogenic signalling role of the ErbB2 originates from the lack of an intramolecular tether in the extracellular region which self-inhibits other ErbB receptors. Several reports showed the immunogenic property of ErbB2 and its potential to produce antibodies in some patients, emphasizing the role of ErbB2 as an interesting candidate in cancer immunotherapy. There is continuous expanding research in the use of plants as tools for the expression of heterologous non-plant origin proteins. These plant-made proteins are often proposed as pharmaceutical molecules, as their production in plants can be safe, quick and cheap with infinite scalability. Plant-made antigens have been already tested in clinical trials and offer the possibility to be administered orally. Here, we investigated the potential of transiently expressing the 631aa-extracellular (EC) region and a portion of the transmembrane (TM) region of rat ErbB2 in Nicotiana benthamiana plants. Rat ErbB2EC5'TM was cloned in fusion with a 5'-terminal His tag in a Cowpea mosaic virus-derived expression vector pEAQ-HT, under the control of the Cauliflower mosaic virus 35S promoter (Sainsbury et al., 2009). The construct was delivered to plants by agroinfiltration, a classical procedure for plant gene expression. The Rat ErbB2EC5'TM was successfully expressed in plants leaves six days post-agroinfiltration. A 73-kDa band of the expected size for Rat ErbB2EC5'TM was detected in Western Blot analyses using either anti-ErbB2 monoclonal antibody and a anti-poly-Histidine reagent. This presents the first report of expression of ErbB2 protein in plants and opens perspectives for further immunological testing plant-produced ErbB2-antigens.
Transient expression of rat ErbB2 protein in Nicotiana benthamiana
Matic S;Noris;
2011
Abstract
The ErbB2 (Her2/neu) gene encodes a 185-kDa glycoprotein that is ligand-less receptor with a tyrosine kinase activity. It is overexpressed in human cancers such as breast, lung, kidney, bladder and ovary carcinomas, while it is low expressed in normal tissues. Structural studies demonstrated that oncogenic signalling role of the ErbB2 originates from the lack of an intramolecular tether in the extracellular region which self-inhibits other ErbB receptors. Several reports showed the immunogenic property of ErbB2 and its potential to produce antibodies in some patients, emphasizing the role of ErbB2 as an interesting candidate in cancer immunotherapy. There is continuous expanding research in the use of plants as tools for the expression of heterologous non-plant origin proteins. These plant-made proteins are often proposed as pharmaceutical molecules, as their production in plants can be safe, quick and cheap with infinite scalability. Plant-made antigens have been already tested in clinical trials and offer the possibility to be administered orally. Here, we investigated the potential of transiently expressing the 631aa-extracellular (EC) region and a portion of the transmembrane (TM) region of rat ErbB2 in Nicotiana benthamiana plants. Rat ErbB2EC5'TM was cloned in fusion with a 5'-terminal His tag in a Cowpea mosaic virus-derived expression vector pEAQ-HT, under the control of the Cauliflower mosaic virus 35S promoter (Sainsbury et al., 2009). The construct was delivered to plants by agroinfiltration, a classical procedure for plant gene expression. The Rat ErbB2EC5'TM was successfully expressed in plants leaves six days post-agroinfiltration. A 73-kDa band of the expected size for Rat ErbB2EC5'TM was detected in Western Blot analyses using either anti-ErbB2 monoclonal antibody and a anti-poly-Histidine reagent. This presents the first report of expression of ErbB2 protein in plants and opens perspectives for further immunological testing plant-produced ErbB2-antigens.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
