Microelectrophoresis and auxilary micromethods

Neuhoff, Volker

doi:10.1002/(sici)1522-2683(20000101)21:1<3::aid-elps3>3.0.co;2-h

Cited by 23 publications

(11 citation statements)

References 90 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PAGE performed under native conditions can also demonstrate aggregation [76,77]. This is inexpensive and can deliver nanogram detection limits with sensitive methods involving staining with, for example, silver (0.2 ng of protein/lane) [78] or Coomassie Brilliant Blue G-250 [79], but they are seriously constrained by size limits, a factor that restricts their usefulness for many biopharmaceuticals. Other problems are the need to exclude SDS and reducing agents, which increase gel running times and prevent analysis of proteins without a net negative charge under native conditions.…”

Section: Increases Aggregationmentioning

confidence: 99%

Strategies for analysing and improving the expression and quality of recombinant proteins made in mammalian cells

Jenkins

Tyther

Murphy

2009

Biotech and App Biochem

View full text Add to dashboard Cite

The production of monoclonal antibodies and other recombinant proteins is one of the highest growth areas in the pharmaceutical industry. Mammalian cells are used to manufacture the majority of biotherapeutics, largely due to their ability to perform complex post-translational modifications. Although significant progress has been made recently in improving product yields and protein quality, many challenges still lie ahead to achieve consistently high yields while avoiding potentially damaging protein modifications. The present review first considers the strategies used to analyse and improve recombinant protein expression of industrial cell lines, with an emphasis on proteomic technologies. Next, cellular and environmental influences on protein production and quality are examined, and strategies for improvements in product yield and quality are reviewed. The analytical techniques required to detect these protein changes are also described, together with prospects for assay improvements.

show abstract

Section: Increases Aggregationmentioning

confidence: 99%

Strategies for analysing and improving the expression and quality of recombinant proteins made in mammalian cells

Jenkins

Tyther

Murphy

2009

Biotech and App Biochem

View full text Add to dashboard Cite

show abstract

“…The second dimension was run on 15% acrylamide gels on a vertical system. Gels were stained with silver nitrate [12] or colloidal CBB [13], or transferred on nitrocellulose membrane for immunoblotting with anti-LTP1 antibodies.…”

Section: -Dementioning

confidence: 99%

Probing heat-stable water-soluble proteins from barley to malt and beer

et al. 2005

View full text Add to dashboard Cite

Proteins determine the quality of barley in malting and brewing end-uses. In this regard, water-soluble barley proteins play a major role in the formation, stability, and texture of head foams. Our objective was to survey the barley seed proteins that could be involved in the foaming properties of beer. Therefore, two-dimensional (2-D) electrophoresis and mass spectrometry were combined to highlight the barley proteins that could resist the heating treatments occurring during malting and brewing processes. As expected, from barley to malt and to beer, most of the heat-stable proteins are disulfide-rich proteins, implicated in the defense of plants against their bio-aggressors, e.g., serpin-like chymotrypsin inhibitors (protein Z), amylase and amylase-protease inhibitors, and lipid transfer proteins (LTP1 and LTP2). For LTP1s, the complex pattern displayed in 2-D electrophoresis could be related to some chemical modifications already described elsewhere, such as acylation or glycation through Maillard reactions, which occur on malting. Our proteomics approach allowed the identification of the numerous proteins present in beer in addition to the major ones already described. The involvement of these proteins in the quality of beer foam can now be evaluated.

show abstract

“…In the past decade, the development of microscale (chipbased) separation devices has experienced an explosive growth in both industrial and academic fields worldwide [1]. Most of the devices reported to date have been fabricated in glass or silicon [2][3][4][5][6].…”

Section: Introductionmentioning

confidence: 99%

A polymeric master replication technology for mass fabrication of poly(dimethylsiloxane) microfluidic devices

Lin

et al. 2005

Electrophoresis

View full text Add to dashboard Cite

A polymeric master replication technology for mass fabrication of poly(dimethylsiloxane) microfluidic devicesA protocol of producing multiple polymeric masters from an original glass master mold has been developed, which enables the production of multiple poly(dimethylsiloxane) (PDMS)-based microfluidic devices in a low-cost and efficient manner. Standard wetetching techniques were used to fabricate an original glass master with negative features, from which more than 50 polymethylmethacrylate (PMMA) positive replica masters were rapidly created using the thermal printing technique. The time to replicate each PMMA master was as short as 20 min. The PMMA replica masters have excellent structural features and could be used to cast PDMS devices for many times. An integration geometry designed for laser-induced fluorescence (LIF) detection, which contains normal deep microfluidic channels and a much deeper optical fiber channel, was successfully transferred into PDMS devices. The positive relief on seven PMMA replica masters is replicated with regard to the negative original glass master, with a depth average variation of 0.89% for 26 mm deep microfluidic channels and 1.16% for the 90 mm deep fiber channel. The imprinted positive relief in PMMA from master-to-master is reproducible with relative standard deviations (RSDs) of 1.06% for the maximum width and 0.46% for depth in terms of the separation channel. The PDMS devices fabricated from the PMMA replica masters were characterized and applied to the separation of a fluorescein isothiocyanate (FITC)-labeled epinephrine sample.

show abstract

Microelectrophoresis and auxilary micromethods

Cited by 23 publications

References 90 publications

Strategies for analysing and improving the expression and quality of recombinant proteins made in mammalian cells

Strategies for analysing and improving the expression and quality of recombinant proteins made in mammalian cells

Probing heat-stable water-soluble proteins from barley to malt and beer

A polymeric master replication technology for mass fabrication of poly(dimethylsiloxane) microfluidic devices

Contact Info

Product

Resources

About