PMID- 25786575
OWN - NLM
STAT- MEDLINE
DCOM- 20170126
LR  - 20170126
IS  - 1470-8744 (Electronic)
IS  - 0885-4513 (Linking)
VI  - 63
IP  - 3
DP  - 2016 May
TI  - Expression of anti-tumor necrosis factor alpha (TNFalpha) single-chain variable 
      fragment (scFv) in Spirodela punctata plants transformed with Agrobacterium 
      tumefaciens.
PG  - 354-61
LID - 10.1002/bab.1373 [doi]
AB  - Therapeutic antibodies against tumor necrosis factor alpha (TNFalpha) have been 
      considered effective for some of the autoimmune diseases such as rheumatoid 
      arthritis, Crohn's diseases, and so on. But associated limitations of the current 
      therapeutics in terms of cost, availability, and immunogenicity have necessitated 
      the need for alternative candidates. Single-chain variable fragment (scFv) can 
      negate the limitations tagged with the anti-TNFalpha therapeutics to a greater 
      extent. In the present study, Spirodela punctata plants were transformed with 
      anti-TNFalpha through in planta transformation using Agrobacterium tumefaciens 
      strain, EHA105. Instead of cefotaxime, garlic extract (1 mg/mL) was used to 
      remove the agrobacterial cells after cocultivation. To the best of our knowledge, 
      this report shows for the first time the application of plant extracts in 
      transgenic plant development. 95% of the plants survived screening under 
      hygromycin. ScFv cDNA integration in the plant genomic DNA was confirmed at the 
      molecular level by PCR. The transgenic protein expression was followed up to 
      10 months. Expression of scFv was confirmed by immunodot blot. Protein expression 
      levels of up to 6.3% of total soluble protein were observed. beta-Glucuronidase and 
      green fluorescent protein expressions were also detected in the antibiotic 
      resistant plants. The paper shows the generation of transgenic Spirodela 
      punctuata plants through in planta transformation.
CI  - (c) 2015 International Union of Biochemistry and Molecular Biology, Inc.
FAU - Balaji, Parthasarathy
AU  - Balaji P
AD  - Centre for BioSeparation Technology (CBST), VIT University, Vellore, Tamil Nadu, 
      632014, India.
FAU - Satheeshkumar, P K
AU  - Satheeshkumar PK
AD  - Centre for BioSeparation Technology (CBST), VIT University, Vellore, Tamil Nadu, 
      632014, India.
FAU - Venkataraman, Krishnan
AU  - Venkataraman K
AD  - Centre for BioSeparation Technology (CBST), VIT University, Vellore, Tamil Nadu, 
      632014, India.
FAU - Vijayalakshmi, M A
AU  - Vijayalakshmi MA
AD  - Centre for BioSeparation Technology (CBST), VIT University, Vellore, Tamil Nadu, 
      632014, India.
LA  - eng
PT  - Journal Article
DEP - 20150704
PL  - United States
TA  - Biotechnol Appl Biochem
JT  - Biotechnology and applied biochemistry
JID - 8609465
RN  - 0 (Single-Chain Antibodies)
RN  - 0 (Tumor Necrosis Factor-alpha)
SB  - IM
MH  - Agrobacterium tumefaciens/*genetics
MH  - Araceae/*genetics
MH  - Escherichia coli/genetics
MH  - Gene Expression
MH  - *Genetic Engineering
MH  - Genetic Vectors/genetics
MH  - Plants, Genetically Modified
MH  - Single-Chain Antibodies/*genetics/*immunology
MH  - *Transformation, Genetic
MH  - Tumor Necrosis Factor-alpha/*immunology
OTO - NOTNLM
OT  - Agrobacterium tumefaciens
OT  - Lemnaceae: Spirodela punctata
OT  - anti-TNF alpha scFv
OT  - anti-agrobacterial plant extracts
OT  - immobilized metal affinity chromatography (IMAC)
OT  - in planta transformation
EDAT- 2015/03/20 06:00
MHDA- 2017/01/27 06:00
CRDT- 2015/03/20 06:00
PHST- 2015/01/29 00:00 [received]
PHST- 2015/03/14 00:00 [accepted]
PHST- 2015/03/20 06:00 [entrez]
PHST- 2015/03/20 06:00 [pubmed]
PHST- 2017/01/27 06:00 [medline]
AID - 10.1002/bab.1373 [doi]
PST - ppublish
SO  - Biotechnol Appl Biochem. 2016 May;63(3):354-61. doi: 10.1002/bab.1373. Epub 2015 
      Jul 4.