Abstract:
:A growing number of studies require the purification of multiple proteins simultaneously and the development of simple economical high-throughput purification methods is essential. We have tested the purification of two related proteins in a variety of conditions to benchmark the semi-automated affinity chromatography method for the QIAcube that we have developed. We find that this new QIAcube method can successfully purify milligram quantities of proteins with minimal user involvement and performs as well as methods based on gravity. The method could easily be adapted to other chromatography resins and should prove to be a versatile method for optimizing protein expression or purification conditions for multiple proteins while obtaining sufficient amounts for subsequent biochemical analyses.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
McGraw J,Tatipelli VK,Feyijinmi O,Traore MC,Eangoor P,Lane S,Stollar EJdoi
10.1016/j.pep.2014.01.014subject
Has Abstractpub_date
2014-04-01 00:00:00pages
48-53eissn
1046-5928issn
1096-0279pii
S1046-5928(14)00016-3journal_volume
96pub_type
杂志文章abstract::In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.09.004
更新日期:2009-01-01 00:00:00
abstract::Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 is 60 amino acids long and lacks one of the three conserved disulfide bonds typically found in Kunitz domain modules. It binds specifically to voltage activated potassium channe...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.01.019
更新日期:2006-06-01 00:00:00
abstract::Human cystatin C (HCC), encoded by cystatin 3 gene, is a 13.3kDa endogenous cysteine proteinase inhibitor and an important biomarker of renal function. However, expressing recombinant cystatin C is difficult because of low yield and inclusion bodies in Escherichia coli (E. coli). In this study, we cloned HCC gene into...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.03.006
更新日期:2015-07-01 00:00:00
abstract::A kappa-light chain from a Fab expression system was truncated by the insertion of a stop codon in the gene sequence to produce a variable light (VL) single domain antibody (dAb). Here, we describe the expression of dAb in the periplasm of Escherichia coli through fermentation in a defined media. Immunoglobulin bindin...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.07.013
更新日期:2007-02-01 00:00:00
abstract::The hexokinase (ATP;D-hexose 6-phosphotransferase, EC 2.7.1.1) of Schistosoma mansoni has been expressed in Escherichia coli as a fusion protein including an N-terminal polyhistidine tag and enterokinase cleavage site. The enzyme was purified by metal chelate chromatography to > 95% homogeneity, based on analysis by S...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0113
更新日期:1996-11-01 00:00:00
abstract::Vascular endothelial growth factor (VEGF) is one of the most significant mediators of angiogenesis, which interacts with a specific membrane receptor: VEGF receptor 2 (VEGFR2). Studies elsewhere have shown that, a VEGF-blocker can regulate several vital processes of tumor promotion. However, there is no literature evi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2013.04.010
更新日期:2013-08-01 00:00:00
abstract::Antimicrobial peptides (AMPs) consist of molecules acting on the defense systems of numerous organisms toward tumor and multiple pathogens, such as bacteria, fungi, viruses, and parasites. Compared to traditional antibiotics, AMPs are more stable and have lower propensity for developing resistance through functioning ...
journal_title:Protein expression and purification
pub_type: 杂志文章,评审
doi:10.1016/j.pep.2017.08.003
更新日期:2017-12-01 00:00:00
abstract::The plant protein production system is a platform that can not only reduce production costs but also produce monoclonal antibodies that do not have the risk of residual proteins from the host. However, due to the difference between post-translational processes in plants and animals, there may be a modification in the ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.03.004
更新日期:2019-07-01 00:00:00
abstract::A family of restriction enzyme- and ligation-independent cloning vectors has been developed for producing recombinant His-tagged fusion proteins in Escherichia coli. These are based on pURI2 and pURI3 expression vectors which have been previously used for the successful production of recombinant proteins at the millig...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2010.10.013
更新日期:2011-03-01 00:00:00
abstract::Malassezia globosa is pathogenic fungus that causes skin disorders including dandruff in humans. Many yeast cytochrome CYP enzymes are involved in the biosynthesis of sterols and are considered major targets of azole antifungal agents. Here, we report on the expression and characterization of the MGL_0310 gene product...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.07.002
更新日期:2015-10-01 00:00:00
abstract::The catalase gene of Psychrobacter sp. T-3 was cloned, and the gene product (PktA) was overexpressed in Escherichia coli. The specific activity of the purified PktA was slightly lower than that of the native purified enzyme obtained from Psychrobacter sp. T-3. Spectrophotometric measurements of the purified enzymes su...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.03.016
更新日期:2008-06-01 00:00:00
abstract::A cysteine-rich (approximately 20%), low molecular weight (MW 6 kDa) polypeptide has been isolated from the Korean blood-sucking leech, Hirudo nipponia. From its amino acid composition and N-terminal amino acid sequence analysis, the new protein is similar to granulin (or epithelin), and so it has been named leech gra...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1077
更新日期:1999-07-01 00:00:00
abstract::High affinity interaction between S-protein and S-peptide fragments of bovine pancreatic RNase A has been recently used for construction of molecular vehicles for targeted drug delivery. The vehicle is assembled as a complex of drug carrier conjugated S-protein with S-peptide-tagged targeting protein. To avoid random ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00546-6
更新日期:2002-12-01 00:00:00
abstract::The XRCC1 DNA repair protein contains two regions of approximately 100 amino acids each that share homology with the BRCT (BRCA1 carboxyl terminus) domain superfamily. These two regions of XRCC1 have been shown to interact independently with DNA ligase III and poly(ADP-ribose)polymerase as part of a mechanism involved...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1070
更新日期:1999-07-01 00:00:00
abstract::Escherichia coli oligoribonuclease (EcoORN), encoded by the orn gene, is a 3'-5' exonuclease that degrades short single-stranded oligoribonucleotides to rNMPs in the final step of RNA degradation. The orn gene is essential in E. coli, but not in higher organisms, and close homologues are present in other genomes from ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.10.005
更新日期:2008-02-01 00:00:00
abstract::Expression of recombinant proteins is an important step towards elucidating the functions of many genes discovered through genomic sequencing projects. It is also critical for validating gene targets and for developing effective therapies for many diseases. Here we describe a novel method to express recombinant protei...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00075-5
更新日期:2003-07-01 00:00:00
abstract::The ELR-CXC chemokines are important to neutrophil inflammation in many acute and chronic diseases. Among them, CXCL8 (interleukin-8, IL-8), binds to both the CXCR1 and CXCR2 receptors with high affinity and the expression levels of CXCL8 are elevated in many inflammatory diseases. Recently, an analogue of human CXCL8...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.04.007
更新日期:2008-09-01 00:00:00
abstract::Chloroplast transformation systems offer unique advantages in biotechnology, including high level of foreign gene expression, maternal inheritance, and polycistronic expression. We studied chloroplast expression of LTK63 (change Ser-->Lys at position 63 in the A subunit) which is the mutant of Escherichia coli heat-la...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.08.002
更新日期:2004-11-01 00:00:00
abstract::We have cloned ansB (YPTB1411) gene from Yersinia pseudotuberculosis Q66CJ2 and constructed stable inducible expression system that overproduce L-asparaginase from Y. pseudotuberculosis (YpA) in Escherichiacoli BL21 (DE3) cells. For purification of YpA we used Q-Sepharose and DEAE-Toyopearl column chromatography. We e...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.12.005
更新日期:2012-03-01 00:00:00
abstract::The transcription factor IIIC (TFIIIC) is a multisubunit DNA-binding factor required for promoter recognition and TFIIIB assembly on tRNA genes transcribed by RNA polymerase III. Yeast TFIIIC consists of six subunits, organized in the two globular subcomplexes tauA and tauB, which recognize two internal tDNA promoter ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.06.013
更新日期:2006-02-01 00:00:00
abstract::Achromogenic atypical Aeromonas salmonicida is the causative agent of goldfish ulcer disease. Virulence of this bacterium is associated with the production of a paracrystalline outer membrane A-layer protein. The species-specific structural gene for the monomeric form of A-protein was cloned into a pET-3d plasmid in o...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1054
更新日期:1999-08-01 00:00:00
abstract::The gene coding for phosphoglucomutase (PGM) from Oryctolagus cuniculus (rabbit) has been expressed in Escherichia coli under a T7 expression system with a His-tag. About half of the expressed PGM protein was present in inclusion bodies, but this protein was inactive when solubilized. The protein in the soluble cell f...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1288
更新日期:2000-10-01 00:00:00
abstract::Triacylglycerol hydrolase mobilizes stored triacylglycerol some of which is used for very-low-density lipoprotein assembly in the liver. A full-length cDNA coding for a human triacylglycerol hydrolase (hTGH) was isolated from a human liver cDNA library. The cDNA has an open reading frame of 576 amino acids with a clea...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1553
更新日期:2002-02-01 00:00:00
abstract::Recombinant cytochrome P450 (CYP or P450) enzymes are useful for drug metabolism research and thereby many expression and purification systems have been developed. Here, we provide a method for the purification of human P450s 3A4 and 1A2 expressed in Escherichia coli using mixed micelles containing anionic phospholipi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.05.010
更新日期:2014-09-01 00:00:00
abstract::Post-translational modification of proteins is a dynamic way of generating new protein-protein interaction interfaces that are critical for signaling networks in diverse cellular functions. Purified recombinant proteins frequently lack these signature modifications. Using the tumor suppressor p53 as the model protein,...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.01.015
更新日期:2005-06-01 00:00:00
abstract::A recombinant Hc fragment of botulinum neurotoxin, serotype B (rBoNTB(Hc)), has been successfully expressed in a Mut+ strain of the methylotrophic yeast Pichia pastoris for use as an antigen in a proposed human vaccine. The fermentation process consisted of batch phase on glycerol, followed by glycerol and methanol fe...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0910
更新日期:1998-08-01 00:00:00
abstract::L1 is a human cell adhesion glycoprotein involved in the development of the central nervous system that comprises six immunoglobulin-like domains (Ig1-Ig6), five fibronectin-type III (FN1-FN5) domains, a single transmembrane region and a cytoplasmic domain. It contains 20 potential N-glycosylation sites and is heavily...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.10.008
更新日期:2007-03-01 00:00:00
abstract::The Gradiflow, a preparative electrophoresis instrument designed to separate molecules on the basis of their size and charge, was used to purify antibody Fab and F(ab')2 fragments. The method described is charge based, utilizing the difference in the pI between the antibody Fab/F(ab')2 fragments and antibody Fc fragme...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/S1046-5928(03)00219-5
更新日期:2003-11-01 00:00:00
abstract::A hexahistidine tag was fused to the N-terminus of apoaequorin. A suitable vector encoding the fusion protein was constructed and used for transformation of Escherichia coli JM109 cells. Apoaequorin was overexpressed under the control of tac promoter. It was found, however, that most of the protein existed in the form...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00614-9
更新日期:2003-02-01 00:00:00
abstract::The Human Secretome Project aims to produce and purify all human secreted proteins as full-length. In order to enable this, a robust, gentle and effective purification process is needed, where multiple proteins can be purified in parallel. For this reason, a purification system based on a Protein C-tag and the HPC4 an...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105698
更新日期:2020-11-01 00:00:00