Cloning, expression and purification of folate receptor alpha precursor from Homo sapiens for bioanalytical applications for cancer research

• Grant number: 11/16364-6
• Support Opportunities: Scholarships in Brazil - Master
• Start date: May 01, 2012
• End date: August 31, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Emanuel Carrilho
• Grantee: Beatriz Nogueira Messias de Miranda
• Host Institution: Instituto de Química de São Carlos (IQSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
• Associated scholarship(s): 13/15892-4 - Production of polymersome vesicles for bioanalytical applications in cancer research, BE.EP.MS

Abstract

In recent years the incidence of cancer has grown significantly in the global context, so that, according to the International Agency for Research on Cancer, the overall impact of this disease more than doubled in 30 years. The techniques currently employed nowadays for the detection of the disease are not individually conclusive because of the subjectivity of the analysis, in addition to being highly invasive, does not allow early diagnosis of disease. Therefore, is generated a motivation for the development of new alternative technologies for the detection of cancer. Recently, Lima et al. (2010) reported the possibility of the use of biosensors for the detection of biomarkers in urine, blood or other bodily fluids, facilitating the diagnosis of disease in terms of operation, cost and speed.Cancer cells have specific receptors with high affinity to folate and are highly replicative, these cells are extremely dependent on the supply of reduced folate. With rare exceptions, alpha folate receptor (FR-±) is induced selectively in cancerous tissues and rarely expressed in normal cells, being thus highly sensitive and specific. FRs-± are extracellular proteins of 239 amino acid residues with specific biochemical properties which can be exploited by marking strategy for detection and treatment of cancer (biomarkers).This project propose cloning, expression and purification of the FR-± for subsequent immobilization of the same in microsystem and studies with the biosensor developed by Lima et al. (2010).Thus, bioanalytical biocaracterização or characterization of recombinant protein FR-± (r-FR-±) is useful for understanding the role of this protein in the development of tumor tissues and development of drugs that potentially inactivate this protein, such as the antifolates , which are now the most investigated anticancer agents.

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MESSIAS MIRANDA, BEATRIZ NOGUEIRA; FOTORAN, WESLEY LUZETTI; CANDURI, FERNANDA; FERREIRA SOUZA, DULCE HELENA; WUNDERLICH, GERHARD; CARRILHO, EMANUEL. Heterologous expression of Homo sapiens alpha-folate receptors in E. coli by fusion with a trigger factor for enhanced solubilization. Protein Expression and Purification, v. 142, p. 75-80, FEB 2018. (11/16364-6, 08/57805-2, 13/15892-4, 09/54040-8)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

MIRANDA, Beatriz Nogueira Messias de. Cloning, expression and purification of alpha folate receptors from Homo sapiens for bioanalitical aplications in cancer research. 2014. Master's Dissertation - Universidade de São Paulo (USP). Instituto de Química de São Carlos (IQSC/BT) São Carlos.