Evidence map›Paper›PMID 25349981›Full record

ArticleJournal of visualized experiments : JoVE2014

Recombinant protein expression for structural biology in HEK 293F suspension cells: a novel and accessible approach.

Nicola Portolano, Peter J Watson, Louise Fairall, Christopher J Millard, Charles P Milano, Yun Song, Shaun M Cowley, John W R Schwabe

Open access · greenAbstract readVideo-Audio Media
In one paragraph

Article in Journal of visualized experiments : JoVE, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.

0numbers the graph read from it
0cells of the map it votes in
47citing papers in PubMed
1.4field-weighted citation impact, top 20% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

47 citing papers in PubMed, 70 citations in OpenAlex.

  1. Article
  2. Article
  3. Article
  4. Article
  5. Article
  6. Article
  7. Article
  8. Article
  9. Article
  10. Review
  11. Article
  12. Article
  13. Article
  14. Article
  15. Article
  16. Article
  17. Article
  18. Article
  19. Article
  20. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 1 institution in 1 country.

Nicola PortolanoDepartment of Biochemistry, University of Leicester.
Peter J WatsonDepartment of Biochemistry, University of Leicester.
Louise FairallDepartment of Biochemistry, University of Leicester.
Christopher J MillardDepartment of Biochemistry, University of Leicester.
Charles P MilanoDepartment of Biochemistry, University of Leicester.
Yun SongDepartment of Biochemistry, University of Leicester.
Shaun M CowleyDepartment of Biochemistry, University of Leicester.
John W R SchwabeDepartment of Biochemistry, University of Leicester; js336@leicester.ac.uk.
University of Leicester · GB

Funding

Biotechnology and Biological Sciences Research Council RM31G0224Medical Research Council G0600135Medical Research Council MR/J009202/1Wellcome Trust 085408Wellcome Trust 100237Wellcome Trust 100237/Z/12/Z
6 · The paper itself

Abstract

The expression and purification of large amounts of recombinant protein complexes is an essential requirement for structural biology studies. For over two decades, prokaryotic expression systems such as E. coli have dominated the scientific literature over costly and less efficient eukaryotic cell lines. Despite the clear advantage in terms of yields and costs of expressing recombinant proteins in bacteria, the absence of specific co-factors, chaperones and post-translational modifications may cause loss of function, mis-folding and can disrupt protein-protein interactions of certain eukaryotic multi-subunit complexes, surface receptors and secreted proteins. The use of mammalian cell expression systems can address these drawbacks since they provide a eukaryotic expression environment. However, low protein yields and high costs of such methods have until recently limited their use for structural biology. Here we describe a simple and accessible method for expressing and purifying milligram quantities of protein by performing transient transfections of suspension grown HEK (Human Embryonic Kidney) 293 F cells.

Indexed as

BiotechnologyHEK293 CellsHumansMolecular BiologyRecombinant ProteinsTransfectionRecombinant Proteins

Identifiers

PMID25349981
PMCPMC4420617
OpenAlexW1980329019

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.