PMID- 26852026
OWN - NLM
STAT- MEDLINE
DCOM- 20170116
LR  - 20221207
IS  - 1559-0291 (Electronic)
IS  - 0273-2289 (Linking)
VI  - 179
IP  - 2
DP  - 2016 May
TI  - Single-Step Purification and Characterization of A Recombinant Serine Proteinase 
      Inhibitor from Transgenic Plants.
PG  - 220-36
LID - 10.1007/s12010-016-1989-8 [doi]
AB  - Expression of recombinant therapeutic proteins in transgenic plants has a 
      tremendous impact on safe and economical production of biomolecules for 
      biopharmaceutical industry. The major limitation in their production is 
      downstream processing of recombinant protein to obtain higher yield and purity of 
      the final product. In this study, a simple and rapid process has been developed 
      for purification of therapeutic recombinant alpha1-proteinase inhibitor (ralpha1-PI) from 
      transgenic tomato plants, which is an abundant serine protease inhibitor in human 
      serum and chiefly inhibits the activity of neutrophil elastase in lungs. We have 
      expressed ralpha1-PI with modified synthetic gene in transgenic tomato plants at a 
      very high level ( approximately 3.2 % of total soluble protein). The heterologous protein was 
      extracted with (NH4)2SO4 precipitation, followed by chromatographic separation on 
      different matrices. However, only immunoaffinity chromatography resulted into 
      homogenous preparation of ralpha1-PI with 54 % recovery. The plant-purified ralpha1-PI 
      showed molecular mass and structural conformation comparable to native serum 
      alpha1-PI, as shown by mass spectrometry and optical spectroscopy. The results of 
      elastase inhibition assay revealed biological activity of the purified ralpha1-PI 
      protein. This work demonstrates a simple and efficient one-step purification of 
      ralpha1-PI from transgenic plants, which is an essential prerequisite for further 
      therapeutic development.
FAU - Jha, Shweta
AU  - Jha S
AD  - Plant Transgenic Lab, Genetics and Molecular Biology Division, CSIR-National 
      Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, Lucknow, 226001, 
      U.P., India. jha.shweta80@gmail.com.
AD  - Department of Botany (Centre of Advanced Study), Jai Narain Vyas University, 
      Jodhpur, 342001, India. jha.shweta80@gmail.com.
FAU - Agarwal, Saurabh
AU  - Agarwal S
AD  - Plant Transgenic Lab, Genetics and Molecular Biology Division, CSIR-National 
      Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, Lucknow, 226001, 
      U.P., India.
AD  - Baylor College of Medicine, Houston, TX, USA.
FAU - Sanyal, Indraneel
AU  - Sanyal I
AD  - Plant Transgenic Lab, Genetics and Molecular Biology Division, CSIR-National 
      Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, Lucknow, 226001, 
      U.P., India.
FAU - Amla, D V
AU  - Amla DV
AD  - Plant Transgenic Lab, Genetics and Molecular Biology Division, CSIR-National 
      Botanical Research Institute, Rana Pratap Marg, P.O. Box 436, Lucknow, 226001, 
      U.P., India.
LA  - eng
PT  - Journal Article
DEP - 20160206
PL  - United States
TA  - Appl Biochem Biotechnol
JT  - Applied biochemistry and biotechnology
JID - 8208561
RN  - 0 (Recombinant Proteins)
RN  - 0 (alpha 1-Antitrypsin)
SB  - IM
MH  - Chromatography, Affinity
MH  - Humans
MH  - Solanum lycopersicum/chemistry/*genetics
MH  - Plants, Genetically Modified/chemistry/genetics
MH  - Protein Conformation
MH  - Recombinant Proteins/biosynthesis/chemistry/genetics/*isolation & purification
MH  - alpha 1-Antitrypsin/*biosynthesis/chemistry/genetics/*isolation & purification
OTO - NOTNLM
OT  - Downstream processing
OT  - Heterologous protein expression
OT  - Molecular pharming
OT  - Protein purification
OT  - Recombinant therapeutic proteins
OT  - Serine proteinase inhibitor
OT  - Transgenic tomato plants
EDAT- 2016/02/08 06:00
MHDA- 2017/01/17 06:00
CRDT- 2016/02/08 06:00
PHST- 2015/10/13 00:00 [received]
PHST- 2016/01/12 00:00 [accepted]
PHST- 2016/02/08 06:00 [entrez]
PHST- 2016/02/08 06:00 [pubmed]
PHST- 2017/01/17 06:00 [medline]
AID - 10.1007/s12010-016-1989-8 [pii]
AID - 10.1007/s12010-016-1989-8 [doi]
PST - ppublish
SO  - Appl Biochem Biotechnol. 2016 May;179(2):220-36. doi: 10.1007/s12010-016-1989-8. 
      Epub 2016 Feb 6.