High Level Expression of Tissue-Nonspecific Alkaline Phosphatase in the Milk of Transgenic Rabbits
- First Online:
Alkaline phosphatase is a promising therapeutic agent in the Gram-negative bacterial lipopolysaccharide (LPS) mediated acute and chronic diseases. Contrary to other alkaline phosphatase isozymes, purified tissue-nonspecific alkaline phosphatase (TNAP) is not available in large quantities from tissue sources, which would enable to analyse its efficacy in animal sepsis models. Two transgenic rabbit lines were created by pronuclear microinjection with the whey acidic protein promoter-humanTNAP minigene (WAP-hTNAP). Lactating females of both lines produced biologically active human TNAP. As indicated by fractionation of milk samples the recombinant alkaline phosphatase was associated with the membrane of milk fat globules. Alkaline phosphatase enzymatic activity was two orders of magnitude higher compared to normal human serum levels. The demonstration that this TNAP is physiologically active would provide the clue to use transgenic animals as bioreactor for bulk production of the human tissue-nonspecific alkaline phosphatase in milk. This may be a valuable and possibly viable option with important implication in attenuating LPS mediated inflammatory responses.
KeywordsTissue-nonspecific alkaline phosphatase LPS Transgenic rabbit milk Membrane associated recombinant protein
Unable to display preview. Download preview PDF.
- Devinoy E, Montoliu L, Baranyi M, Thépot D, Hiripi L, Fontaine M-L, Bodrogi L, Bősze Zs (2005) Analysis of the efficiency of the rabbit whey acidic protein gene 5′ flanking region in controlling the expression of homologous and heterologous linked genes. J Dairy Res 72:113–119PubMedCrossRefGoogle Scholar
- Poelstra K, Bakker WW, Klok PA, Hardonk MJ, Meijer DK (1997a) A physiologic function for alkaline phosphatase: endotoxin detoxification. Lab Invest 76:319–327Google Scholar
- Poelstra K, Bakker WW, Klok PA, Kamps JA, Hardonk MJ, Meijer DK (1997b) Dephosphorylation of endotoxin by alkaline phosphatase in vivo. Am J Pathol 151:1163–1169Google Scholar
- Stromme JH, Rustad P, Steensland H, Theodorsen L, Urdal P (2004) Reference intervals for eight enzymes in blood of adult females and males measured in accordance with the International Federation of Clinical Chemistry reference system at 37°C: part of the Nordic Reference Interval Project. Scand J Clin Lab Invest 64:371–384PubMedCrossRefGoogle Scholar