ArticleAdvanced pharmaceutical bulletin2015
Construction and Characterization of Recombinant HEK Cell Over Expressing α4 Integrin.
Article in Advanced pharmaceutical bulletin, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
What it found
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Who cites it
5 citing papers in PubMed, 8 citations in OpenAlex.
- Generation of HBsAg DNA aptamer using modified cell-based SELEX strategy.Molecular biology reports · 2021Article
- Selection and characterization of single-stranded DNA aptamers against interleukin-5.Research in pharmaceutical sciences · 2019Article
- Display of human and rabbit monocyte chemoattractant protein-1 on human embryonic kidney 293T cell surface.Research in pharmaceutical sciences · 2018Article
- Construction and characterization of human embryonic kidney-(HEK)-293T cell overexpressing truncated α4 integrin.Research in pharmaceutical sciences · 2018Article
- The silencing effect of miR-30a onResearch in pharmaceutical sciences · 2017Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeIntegrins 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.
methodsEscherichia coli strain top 10, was used as host for ITGA-4 expression vector encoding the human integrin α4. 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.
resultsFlow cytometric analysis showed that HEK-293 cells have no expression of integrin α4 on their surface while 95% of transfected HEK-293 cells with ITGA4, expressed different levels of integrin α4 on their surfaces which correlates well with genomic DNA PCR amplification results.
conclusionThe results suggest that we have successfully constructed the integrin α4 expressing HEK293 cell, which will facilitate further research into the production of antibody, nanobody and aptamer against α4 integrin.
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