Expression of active recombinant human lactoferrin in the milk of transgenic goats.
This report details the establishment of a transgenic goat model in order to produce human lactoferrin (hLf) in the mammary gland for large-scale application and research. Two transgenic male goats were generated by microinjecting sequence encoding hLf cDNA to the pronuclear. In the two lines, derived from the two founders, eight lactating female goats could secrete recombinant human lactoferrin (rhLf) at concentrations of up to 0.765 mg/ml. The method of purifying the rhLf from the milk was achieved using ion-exchange chromatography and resulted in 97% purity. Biochemical and physicochemical characteristics of rhLf were similar to native lactoferrin (nhLf); this included N-terminal sequence, isoelectric point, molecular mass, glycosylation, iron-binding/releasing ability, thermal stability, and proteolysis. The rhLf showed broad spectrum antibacterial activity inhibiting the growth of several pathogenic bacterial strains. Also investigated, although to a lesser degree, was a practicable pasteurization method for the downstream processing of rhLf and, further, a method for the oral administration of rhLf. On the basis of these results, our studies show an optimistic and promising approach for the large-scale production and therapeutic application of rhLf expressed in transgenic goats.
Zhang
Jie
J
Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Li
Lin
L
Cai
Yinfeng
Y
Xu
Xujun
X
Chen
Juan
J
Wu
Youbing
Y
Yu
Huiqing
H
Yu
Guohua
G
Liu
Siguo
S
Zhang
Aimin
A
Chen
Jianquan
J
Cheng
Guoxiang
G
D000818:Animals; D030801:Animals, Genetically Modified; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D015139:Blotting, Southern; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006031:Glycosylation / Q000187:drug effects; D006041:Goats / Q000235:genetics*; D006801:Humans; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008892:Milk / Q000378:metabolism*; D010447:Peptide Hydrolases / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism*; D013696:Temperature; D013816:Thermodynamics; D019076:Transgenes
D000900:Anti-Bacterial Agents; D011994:Recombinant Proteins; D007501:Iron; D010447:Peptide Hydrolases; D007781:Lactoferrin