PMID- 28878310
OWN - NLM
STAT- MEDLINE
DCOM- 20190513
LR  - 20220409
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 7
IP  - 1
DP  - 2017 Sep 6
TI  - Large-scale production of recombinant human lactoferrin from high-expression, 
      marker-free transgenic cloned cows.
PG  - 10733
LID - 10.1038/s41598-017-11462-z [doi]
LID - 10733
AB  - Human lactoferrin (hLF) is a valuable protein for pharmaceutical products and 
      functional foods, and worldwide demand for this protein has steadily increased. 
      However, large-scale recombinant human lactoferrin (rhLF) production using 
      current animal bioreactor techniques is limited by the low expression of foreign 
      proteins, the use of antibiotic resistance genes and the down-regulation of 
      endogenous milk proteins. Here, we generated a herd of marker-free, hLF bacterial 
      artificial chromosome (BAC) transgenic cloned cows, as confirmed by Polymerase 
      chain reaction, Southern blot and Western blot analyses. These transgenic cloned 
      cows produced rhLF in milk at concentrations of 4.5-13.6 g/L. Moreover, the total 
      protein content of the milk was increased. Over two hundred transgenic cloned 
      cows were propagated by multiple ovulation and embryo transfer (MOET). A total of 
      400-450 g of rhLF protein, which shows similar enzymatic activity to natural hLF 
      in iron binding and release, can be purified on a large scale from >100 L of milk 
      per day. Our results suggested that transgenic bovine mammary bioreactors have 
      the potential for large-scale protein production.
FAU - Wang, Ming
AU  - Wang M
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China.
FAU - Sun, Zhaolin
AU  - Sun Z
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China.
FAU - Yu, Tian
AU  - Yu T
AD  - Kejienuo Biotechnology Company, Wuxi, China.
FAU - Ding, Fangrong
AU  - Ding F
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China.
FAU - Li, Ling
AU  - Li L
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China.
FAU - Wang, Xi
AU  - Wang X
AD  - Kejienuo Biotechnology Company, Wuxi, China.
FAU - Fu, Mingbo
AU  - Fu M
AD  - Kejienuo Biotechnology Company, Wuxi, China.
FAU - Wang, Haiping
AU  - Wang H
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China.
FAU - Huang, Jinming
AU  - Huang J
AD  - Dairy cattle Research Center, Academy of Agricultural Sciences, Shandong, China.
FAU - Li, Ning
AU  - Li N
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China. ninglcau@cau.edu.cn.
FAU - Dai, Yunping
AU  - Dai Y
AD  - State Key Laboratory for Agrobiotechnology, College of Biological Sciences, China 
      Agricultural University, Beijing, China. daiyunping@sina.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20170906
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (LTF protein, human)
RN  - 0 (Milk Proteins)
RN  - 0 (Recombinant Proteins)
RN  - E1UOL152H7 (Iron)
RN  - EC 3.4.21.- (Lactoferrin)
SB  - IM
MH  - Animals
MH  - Animals, Genetically Modified
MH  - Bioreactors
MH  - Cattle
MH  - *Gene Expression
MH  - Gene Order
MH  - Genetic Vectors
MH  - Humans
MH  - Iron/metabolism
MH  - Lactoferrin/*biosynthesis/*genetics/isolation & purification
MH  - Milk/metabolism
MH  - Milk Proteins/genetics/metabolism
MH  - Protein Binding
MH  - Recombinant Proteins/*biosynthesis/*genetics/isolation & purification
MH  - Transfection
PMC - PMC5587717
COIS- The authors declare that they have no competing interests.
EDAT- 2017/09/08 06:00
MHDA- 2019/05/14 06:00
PMCR- 2017/09/06
CRDT- 2017/09/08 06:00
PHST- 2017/03/15 00:00 [received]
PHST- 2017/08/25 00:00 [accepted]
PHST- 2017/09/08 06:00 [entrez]
PHST- 2017/09/08 06:00 [pubmed]
PHST- 2019/05/14 06:00 [medline]
PHST- 2017/09/06 00:00 [pmc-release]
AID - 10.1038/s41598-017-11462-z [pii]
AID - 11462 [pii]
AID - 10.1038/s41598-017-11462-z [doi]
PST - epublish
SO  - Sci Rep. 2017 Sep 6;7(1):10733. doi: 10.1038/s41598-017-11462-z.