Sánchez Oliberto, Toledo Jorge R, Rodríguez María P, Castro F O
Department of Transgenesis and Animal Cloning, Animal Biotechnology Division, Centro de Ingeniería Genética y Biotecnología, P.O. Box 6162, Havana 10600, Cuba.
J Biotechnol. 2004 Oct 19;114(1-2):89-97. doi: 10.1016/j.jbiotec.2004.06.009.
The production of large quantities of complex proteins with biopharmaceutical purposes is the main drawback for their more extensive use. Here we demonstrated that a direct instillation of a recombinant adenoviral vector containing an expression cassette for the human growth hormone gene into the mammary gland of mice and goats allowed for the efficient secretion of human growth hormone in the milk. Through this approach we were able to express human growth hormone at maximal levels of 2.8 mg/ml in the milk of mice and up to 0.3 mg/ml in goat milk. We found that the expression levels were closely dependent on both the degree of differentiation of the secretory epithelium and on the adenoviral dose used. Here we demonstrated that the direct transduction of mammary epithelial cells by means of a recombinant adenovirus could be a suitable alternative to transgenic technology for the production of recombinant proteins of biopharmaceutical interest.
出于生物制药目的生产大量复杂蛋白质是其更广泛应用的主要障碍。在此，我们证明将携带人生长激素基因表达盒的重组腺病毒载体直接注入小鼠和山羊的乳腺，可使牛奶中高效分泌人生长激素。通过这种方法，我们能够在小鼠乳汁中以最高2.8毫克/毫升的水平表达人生长激素，在山羊奶中可达0.3毫克/毫升。我们发现表达水平紧密依赖于分泌上皮的分化程度以及所用腺病毒的剂量。在此我们证明，利用重组腺病毒直接转导乳腺上皮细胞可能是一种适用于生产具有生物制药意义的重组蛋白的转基因技术替代方法。
