Expression of active recombinant human lactoferrin in the milk of transgenic goats |
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Authors: | Zhang Jie Li Lin Cai Yinfeng Xu Xujun Chen Juan Wu Youbing Yu Huiqing Yu Guohua Liu Siguo Zhang Aimin Chen Jianquan Cheng Guoxiang |
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Affiliation: | Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China. |
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Abstract: | 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. |
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Keywords: | Human lactoferrin Transgenic goat Protein purification Protein expression Microinjection |
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