| Autor: |
Geada D; Tobacco Research Institute, Tumbadero Road 8.5 km, San Antonio de los Baños, Havana, La Havana, Cuba. dgeadalop@yahoo.com.mx, Valdés R, Escobar A, Ares DM, Torres E, Blanco R, Ferro W, Dorta D, González M, Alemán MR, Padilla S, Gómez L, Del Castillo N, Mendoza O, Urquiza D, Soria Y, Brito J, Leyva A, Borroto C, Gavilondo JV |
| Jazyk: |
angličtina |
| Zdroj: |
Biologicals : journal of the International Association of Biological Standardization [Biologicals] 2007 Oct; Vol. 35 (4), pp. 309-15. Date of Electronic Publication: 2007 Jul 19. |
| DOI: |
10.1016/j.biologicals.2007.02.007 |
| Abstrakt: |
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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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