Expression of human alpha 1 antitrypsin in transgenic sheep.
We have recently described the production of large amounts (< or = 65 grams per litre) of enzymatically active human alpha 1 antitrypsin in the milk of transgenic sheep (Wright et al., 1991). Here, we describe in more detail the expression of the human protein in the milk of these animals throughout the lactation period. Human alpha 1 antitrypsin is also found at much lower levels in the plasma of transgenic ewes before, during and after lactation. It is also detected in male plasma at very low levels. We have previously shown human alpha 1 antitrypsin purified from transgenic sheep milk to be indistinguishable from commercially available human plasma derived alpha 1 antitrypsin in terms of gross sugar content and in vitro activity. Here we extend this comparison to more detailed analyses of glycosylation state, amino-terminal sequence, pI value, and molecular weight determination by mass spectrometry.
Carver
A
A
Pharmaceutical Proteins Ltd., Edinburgh, UK.
Wright
G
G
Cottom
D
D
Cooper
J
J
Dalrymple
M
M
Temperley
S
S
Udell
M
M
Reeves
D
D
Percy
J
J
Scott
A
A
D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D002241:Carbohydrates / Q000737:chemistry; D005260:Female; D006031:Glycosylation; D006801:Humans; D007525:Isoelectric Focusing; D007774:Lactation / Q000378:metabolism*; D013058:Mass Spectrometry; D008892:Milk / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012756:Sheep; D000515:alpha 1-Antitrypsin / Q000032:analysis / Q000096:biosynthesis*
D002241:Carbohydrates; D000515:alpha 1-Antitrypsin