Protein profile and alpha-lactalbumin concentration in the milk of standard and transgenic goats expressing recombinant human butyrylcholinesterase.
The expression of recombinant proteins of pharmaceutical interest in the milk of transgenic farm animals can result in phenotypes exhibiting compromised lactation performance, as a result of the extraordinary demand placed on the mammary gland. In this study, we investigated differences in the protein composition of milk from control and transgenic goats expressing recombinant human butyrylcholinesterase. In Experiment 1, the milk was characterized by gel electrophoresis and liquid chromatography/mass spectrometry in order to identify protein bands that were uniquely visible in the transgenic milk and/or at differing band densities compared with controls. Differences in protein content were additionally evaluated by computer assisted band densitometry. Proteins identified in the transgenic milk only included serum proteins (i.e. complement component 3b, ceruloplasmin), a cytoskeleton protein (i.e. actin) and a stress-induced protein (94 kDA glucose-regulated protein). Proteins exhibiting evident differences in band density between the transgenic and control groups included immunoglobulins, serum albumin, beta-lactoglobulin and alpha-lactalbumin. These results were found to be indicative of compromised epithelial tight junctions, premature mammary cell death, and protein synthesis stress resulting from transgene expression. In Experiment 2, the concentration of alpha-lactalbumin was determined using the IDRing assay and was found to be significantly reduced on day 1 of lactation in transgenic goats (4.33 +/- 0.97 vs. 2.24 +/- 0.25 mg/ml, P < 0.01), but was not different from non-transgenic controls by day 30 (0.99 +/- 0.46 vs. 0.90 +/- 0.11 mg/ml, P > 0.05). We concluded that a decreased/delayed expression of the alpha-lactalbumin gene may be the cause for the delayed start of milk production observed in this herd of transgenic goats.
Baldassarre
H
H
Pharmathene Canada Inc., P.O. Box 320, St. Telesphore, QC, H0P 1Y0, Canada. hernan.baldassarre@pharmathene.com
Schirm
M
M
Deslauriers
J
J
Turcotte
C
C
Bordignon
V
V
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D002091:Butyrylcholinesterase / Q000096:biosynthesis* / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006041:Goats / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007768:Lactalbumin / Q000032:analysis / Q000235:genetics; D007774:Lactation / Q000235:genetics / Q000378:metabolism*; D008892:Milk / Q000737:chemistry / Q000378:metabolism*; D008894:Milk Proteins / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
D008894:Milk Proteins; D011994:Recombinant Proteins; D007768:Lactalbumin; D002091:Butyrylcholinesterase