ABSTRACT
In Biotechnology, the expression of recombinant proteins is a constantly growing field and different hosts are used for this purpose. Some valuable proteins cannot be produced using traditional systems. Insects from the order Lepidoptera infected with recombinant baculovirus have appeared as a good choice to express high levels of proteins, especially those with post-translational modifications. Lepidopteran insects, which are extensively distributed in the world, can be used as small protein factories, the new biofactories. Species like Bombyx mori (silkworm) have been analyzed in Asian countries to produce a great number of recombinant proteins for use in basic and applied science and industry. Many proteins expressed in this larva have been commercialized. Several recombinant proteins produced in silkworms have already been commercialized. On the other hand, species like Spodoptera frugiperda, Heliothis virescens, Rachiplusia nu, Helicoverpa zea and Trichoplusia ni are widely distributed in both the occidental world and Europe. The expression of recombinant proteins in larvae has the advantage of its low cost in comparison with insect cell cultures. A wide variety of recombinant proteins, including enzymes, hormones and vaccines, have been efficiently expressed with intact biological activity. The expression of pharmaceutically proteins, using insect larvae or cocoons, has become very attractive. This review describes the use of insect larvae as an alternative to produce commercial recombinant proteins.
Subject(s)
Insecta/metabolism , Recombinant Proteins/biosynthesis , Animals , Baculoviridae/genetics , Humans , Insecta/genetics , Larva/genetics , Larva/metabolism , Protein Processing, Post-Translational , Recombinant Proteins/geneticsABSTRACT
The immunogenicity and cytotoxicity associated with early generations of adenoviral vectors provided a strong incentive for the development of helper-dependent adenovirus, a last generation of adenoviral vectors that is devoid of all viral coding sequences. These vectors have shown to mediate longer high-level transgene expression in vivo with reduced toxicity and thus offer enormous potential for human gene therapy. In addition, they possess a considerably larger cloning capacity than conventional adenoviral vectors making the transfer of large cDNAs, multiple transgenes and longer tissue-specific or regulable promoters possible. In this article, we review the progress made with helper-dependent adenoviral vectors. The development and optimization of scalable production processes and strategies for helper removal will be presented. Current chromatography options available for vector purification and the new challenges facing researchers for the separation of empty particles and/or helper viruses will be discussed. Finally, we will describe recent advances made in our understanding of their interaction with the immune system and their potential as gene delivery vehicles in vivo for the treatment of diseases affecting liver, skeletal muscle and brain.
Subject(s)
Adenoviridae/genetics , Genetic Therapy , Genetic Vectors , Helper Viruses/genetics , Animals , HumansABSTRACT
El síndrome de Sjogren (SS) es un desorden crónico de tipo inflamatorio autoinmune que compromete las glándulas salivares y oculares. Debido a que sólo se realizan prueban indirectas que midan la funcionalidad del ojo se propuso estandarizar la técnica de biopsia conjuntival en el estudio de los pacientes con síndrome de Sjogren (SS) por medio de un estudio descriptivo y de concordancia en el Hospital Universitario de San Ignacio; se concluyeron 17 voluntarios sanos, 13 pacientes con diagnóstico de ojo seco y 13 pacientes con diagnóstico de SS por los criterios preliminares para clasificación de SS de 1993. Se realizó valoración oftalmológica con tiempo de ruptura de película lagrimal, Schirmer y biopsia conjuntival. Las muestras fueron leídas por 2 patólogos en forma ciega por duplicado. El principal hallazgo histopatológico útil en la diferenciación de la población sana y enferma es la hipoplasia de las células caliciformes (p = 0.054). La arquitectura acinar y ductal e infiltrado linfoplasmocitario no fueron estadísticamente significativas (p = 0.19). En cuanto al estudio de concordancia, para la variación intraobservador para el patólogo A el kappa obtenido mayor de 0.726 y para el patólogo B, mayor de 0.799 indica que la concordancia es buena. La variación intraobservador tiene un kappa aceptable, pero no es el ideal. Por lo que consideramos que la biopsia conjuntival se podría utilizar en el estudio de los pacientes con SS, pero es necesario ampliar el tamaño de la muestra para establecer con mayor exactitud las diferencias entre sanos y enfermos
