ABSTRACT
Antibodies have been one of the proteins widely expressed in tobacco plants for pharmaceutical purposes, which demand contaminant free preparations. Rubisco constitutes 40-60% of tobacco leaf soluble proteins; therefore it is the major potential protein contaminant of plantibodies, while mycotoxins are toxic compounds that could be introduced during the biomass production and post-harvest stages with important consequences to human health. The objective of this paper was to investigate whether Rubisco and mycotoxins are present in Plantibody HB-01 preparations used in the immunopurification of the hepatitis B surface antigen. Rubisco was purified from Nicotiana tabacum yielding 154 microg of protein per gram of leaves and purity over 95%. Among mouse monoclonal antibodies generated against this enzyme, the CBSS.Rub-2 was selected for its immunodetection. It recognizes a conserved sequential epitope of Rubisco large subunit with an affinity constant of 0.13 x 10(8)M(-1). Rubisco quantification limit was 1 microg spreading to the measurement of this contaminant less than 4% of plantibodies samples. Additionally, according to a Reverse Phase-HPLC used to measure the level of adventitiously introduced contaminants, it can be concluded that aflatoxins B1, B2, G1 and G2 were undetected in the purified Plantibody HB-01 samples.
Subject(s)
Aflatoxins/analysis , Hepatitis B Antibodies/isolation & purification , Plantibodies/isolation & purification , Ribulose-Bisphosphate Carboxylase/analysis , Aflatoxins/chemistry , Animals , Antibodies, Monoclonal/genetics , Antibodies, Monoclonal/isolation & purification , Blotting, Western , Chromatography, High Pressure Liquid/methods , Chromatography, High Pressure Liquid/statistics & numerical data , Drug Contamination , Enzyme-Linked Immunosorbent Assay , Hepatitis B Antibodies/genetics , Hepatitis B Surface Antigens/immunology , Humans , Mice , Plantibodies/genetics , Plants, Genetically Modified , Nicotiana/genetics , Nicotiana/immunologyABSTRACT
Se determinaron los niveles de TAR en diferentes marcas de cigarrillos cubanos, tanto de consumo nacional como de exportación en las variedades rubio y negro, con y sin filtro. Se encontraron niveles promedio entre 18,9 y 21,7 mg/cigarrillo de TAR en las variedades rubio sin filtro y de 12,5-12,9 mg en las variedades rubio con filtro. En las variedades de cigarrillos negros, los niveles fueron de 20,1 mg en los cigarrillos sin filtro y entre 18,7-21,3 mg en las variedades de cigarrillo negro con filtro. Los niveles de TAR promedio encontrados en nuestros cigarrillos son comparables con los informados internacionalmente
Subject(s)
Neoplasms , Smoke/analysis , NicotianaABSTRACT
Se determinaron los niveles de TAR en diferentes marcas de cigarrillos cubanos, tanto de consumo nacional como de exportación en las variedades rubio y negro, con y sin filtro. Se encontraron niveles promedio entre 18,9 y 21,7 mg/cigarrillo de TAR en las variedades rubio sin filtro y de 12,5-12,9 mg en las variedades rubio con filtro. En las variedades de cigarrillos negros, los niveles fueron de 20,1 mg en los cigarrillos sin filtro y entre 18,7-21,3 mg en las variedades de cigarrillo negro con filtro. Los niveles de TAR promedio encontrados en nuestros cigarrillos son comparables con los informados internacionalmente
