Production of human monoclonal antibody in eggs of chimeric chickens

Abstract

The tubular gland of the chicken oviduct is an attractive system for protein expression as large quantities of proteins are deposited in the egg, the production of eggs is easily scalable and good manufacturing practices for therapeutics from eggs have been established. Here we examined the ability of upstream and downstream DNA sequences of ovalbumin, a protein produced exclusively in very high quantities in chicken egg white, to drive tissue-specific expression of human mAb in chicken eggs. To accommodate these large regulatory regions, we established and transfected lines of chicken embryonic stem (cES) cells and formed chimeras that express mAb from cES cell-derived tubular gland cells. Eggs from high-grade chimeras contained up to 3 mg of mAb that possesses enhanced antibody-dependent cellular cytotoxicity (ADCC), nonantigenic glycosylation, acceptable half-life, excellent antigen recognition and good rates of internalization.

Manufacturing antibodies in the plant cell

“…All these platforms have dramatically improved their performance in recent years to cope with to the expected shortage of recombinant protein manufacturing capacity [2,3]. For instance, the modifications introduced in prokaryotic systems have made them increasingly able to produce and display complex proteins [4], or have facilitated the introduction of whole metabolic pathways [5]; similarly, yeast strains have been engineered to display nearlyhuman glycosylation profiles [6]; CHO cells have multiplied their yields, reaching g/L levels [7]; alternative animal systems as chicken eggs have arisen [8], and baculovirusbased systems have been adapted to the production of multi-subunit protein complexes [9].…”

Preparation of recombinant proteins in milk to improve human and animal health

“…More than 3000 genes have been knocked out or knocked in this species. The same appears impossible in other species including birds [3]. It was shown that the repeated failure was due to the fact that no genuine pluripotent ES cell lines can be obtained from animals other than two breeds of mice.…”

“…For years, due to the inefficiency of available methods to generate transgenic chickens, no angible success was achieved. Recent studies have shown that ES and PGG cell lines can be implemented to generate transgenic birds [3][4][5] and a monoclonal antibody was produced in egg whites [3]. These experimenters incline to consider that egg white is now becoming a source of pharmaceutical proteins.…”

Spermatogonial Stem Cells and Animal Transgenesis

“…This last application is especially important for species in which embryonic stem cell lines have not been established and other transgenic techniques present limited efficiency. Transgenic animals have huge applications from basic science such as the creation of animal models for human diseases, like Parkinson´s (Crabtree & Zhang, 2011) to production of recombinant pharmaceutic proteins in the animal's fluid: blood, milk (Houdebine, 2000a,b andHoudebine, 2002), egg white (Zhu et al, 2005;van de Lavoir et al, 2006 andLillico et al, 2007) and seminal plasma (Dyck et al, 2003). Ever since the generation of the first transgenic animal, in 1980, through pronuclei microinjection in an embryo's pronuclei (Houdebine, 2009), this method has been used in other prolific species as rat, rabbit and pig (Houdebine, 2000a,b).…”