Production of recombinant albumin by a herd of cloned transgenic cattle

Abstract

Purified plasma derived human albumin has been available as a therapeutic product since World War II. However, cost effective recombinant production of albumin has been challenging due to the amount needed and the complex folding pattern of the protein. In an effort to provide an abundant source of recombinant albumin, a herd of transgenic cows expressing high levels of rhA in their milk was generated. Expression cassettes efficiently targeting the secretion of human albumin to the lactating mammary gland were obtained and tested in transgenic mice. A high expressing transgene was transfected in primary bovine cell lines to produce karyoplasts for use in a somatic cell nuclear transfer program. Founder transgenic cows were produced from four independent cell lines. Expression levels varying from 1-2 g/l to more than 40 g/l of correctly folded albumin were observed. The animals expressing the highest levels of rhA exhibited shortened lactation whereas cows yielding 1-2 g/l had normal milk production. This herd of transgenic cattle is an easily scalable and well characterized source of rhA for biomedical uses.

Non-conventional expression systems for the production of vaccine proteins and immunotherapeutic molecules

“…70 The production of human recombinant albumin, for which the annual need is estimated to be several tons, may be well served by the use of large transgenic animals, replacing the extraction of the protein from human blood. 71 Gaucher disease, an orphan pathology characterized by the inability of the human glucocerebrosidase enzyme to catalyze the breakdown of glucosylceramide, is a low-volume niche market where transgenic animal-derived treatments may have utility. 72 While there are numerous examples of therapeutic molecule production in transgenic livestock, development of vaccine antigen production in these systems appears to be a relatively neglected field of study.…”

Enrichment behavior of immunoglobulin by foam fractionation using response surface methodology

“…Recombinant proteins produced from milk of transgenic dairy animals could be an abundant and pathogenic-free source for immunoglobulin production. Transgenic animals have great capacity to produce large amounts of recombinant proteins at approximately 40 g/L of bulk solution [BOSZE, et al 2008, ECHELARD, et al 2006, ECHELARD, et al 2008, POLLOCK, et al 1999]. This concentration is 50 to 100 times higher than that of conventional bioreactor production methods (0.5-0.8 g/L).…”

Transgenic bovine as bioreactors: Challenges and perspectives

“…The methods investigated include recloning, autologous SCNT, impeding Xist expression from the active X chromosome, treatment with trichostatin A after SCNT and histone deacetylase inhibitors, among others. SCNT has been successfully used for the generation of transgenic cattle for recombinant protein expression in milk aimed at nutritional 12,14 and therapeutic 50,51 purposes. Effective SCNT can be applied to the production of transgenic animals that could contribute toward the development of sustainable and diversified animal production systems.…”