PMID- 26504766
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20151027
LR  - 20200930
IS  - 2228-5881 (Print)
IS  - 2251-7308 (Electronic)
IS  - 2228-5881 (Linking)
VI  - 5
IP  - 3
DP  - 2015 Sep
TI  - Construction and Characterization of Recombinant HEK Cell Over Expressing alpha4 
      Integrin.
PG  - 429-34
LID - 10.15171/apb.2015.058 [doi]
AB  - PURPOSE: Integrins are heterodimeric membrane proteins, which are exposed to post 
      translational modifications in eukaryotic cells in contrast to prokaryotic cells. 
      These modifications provide advantages for production of proper nanobody, mono 
      and polyclonal antibody against this surface protein and also in aptamer 
      selection process. Since the majority of diagnostic and therapeutic antibodies, 
      target the surface epitopes, eukaryotic membrane proteins provide an appropriate 
      model for further investigation on therapeutic agents. METHODS: Escherichia coli 
      strain top 10, was used as host for ITGA-4 expression vector encoding the human 
      integrin alpha4. The plasmid was extracted and consequently, ITGA-4 vector was 
      digested to make a linear plasmid. Human Embryonic Kidney-293 (HEK-293) cell 
      transfected with linear plasmid and subsequently screened for stable ITGA-4 
      expressing Cells. Three separated clones were isolated twenty one days after 
      transfection. Chromosomal DNA was extracted from ITGA-4-transfected cells. The 
      presence of ITGA-4 gene in HEK-293 genome was confirmed by PCR. The expression 
      level of ITGA-4 on HEK-293 cells was also analyzed by Flow cytometry. RESULTS: 
      Flow cytometric analysis showed that HEK-293 cells have no expression of integrin 
      alpha4 on their surface while 95% of transfected HEK-293 cells with ITGA4, expressed 
      different levels of integrin alpha4 on their surfaces which correlates well with 
      genomic DNA PCR amplification results. CONCLUSION: The results suggest that we 
      have successfully constructed the integrin alpha4 expressing HEK293 cell, which will 
      facilitate further research into the production of antibody, nanobody and aptamer 
      against alpha4 integrin.
FAU - Naderi Beni, Shamsi
AU  - Naderi Beni S
AD  - Isfahan University of Medical Sciences, School of Medicine, Department of Genetic 
      and Molecular Biology, Isfahan, Iran.
FAU - Kouhpayeh, Shirin
AU  - Kouhpayeh S
AD  - Isfahan University of Medical Sciences, School of Medicine, Department of 
      Immunology, Isfahan, Iran.
FAU - Hejazi, Zahra
AU  - Hejazi Z
AD  - Isfahan University of Medical Sciences, School of Medicine, Department of Genetic 
      and Molecular Biology, Isfahan, Iran.
FAU - Heidari Hafshejani, Nahid
AU  - Heidari Hafshejani N
AD  - Isfahan University of Medical Sciences, School of Medicine, Department of Genetic 
      and Molecular Biology, Isfahan, Iran.
FAU - Khanahmad, Hossein
AU  - Khanahmad H
AD  - Isfahan University of Medical Sciences, School of Medicine, Department of Genetic 
      and Molecular Biology, Isfahan, Iran.
LA  - eng
PT  - Journal Article
DEP - 20150919
PL  - Iran
TA  - Adv Pharm Bull
JT  - Advanced pharmaceutical bulletin
JID - 101578021
PMC - PMC4616894
OTO - NOTNLM
OT  - Aptamer
OT  - Integrin alpha4
OT  - Multiple Sclerosis
OT  - Natalizumab
OT  - VCAM-1
EDAT- 2015/10/28 06:00
MHDA- 2015/10/28 06:01
PMCR- 2015/09/01
CRDT- 2015/10/28 06:00
PHST- 2014/08/07 00:00 [received]
PHST- 2015/05/13 00:00 [revised]
PHST- 2015/05/14 00:00 [accepted]
PHST- 2015/10/28 06:00 [entrez]
PHST- 2015/10/28 06:00 [pubmed]
PHST- 2015/10/28 06:01 [medline]
PHST- 2015/09/01 00:00 [pmc-release]
AID - 10.15171/apb.2015.058 [doi]
PST - ppublish
SO  - Adv Pharm Bull. 2015 Sep;5(3):429-34. doi: 10.15171/apb.2015.058. Epub 2015 Sep 
      19.