Abstract：
:A tremendous increase has taken place over the last decades in the biochemical and clinical use of antibodies. Unfortunately, the constantly growing demand has not been matched by a corresponding easy access to pure immunoglobulin, as purification procedures tend to be either laborious, expensive, or inefficient. We present a new and simplified method to obtain pure antibody based on the special thermal properties of the streptococcal M proteins, a family of cell-surface exposed coiled-coil molecules which bind different sets of host plasma proteins. The coiled-coil structure is already destabilized at low temperatures and the M proteins unfold reversibly, usually below 40 degrees C. We demonstrate the use of this property to purify immunoglobulin G from rabbit serum with protein H from the AP1 strain of Streptococcus pyogenes. Recombinant protein H is linked to nickel-agarose via a C-terminal histidine tag. After mixing with rabbit serum and washing at room temperature, pure IgG can be eluted from the gel with a moderately heated buffer. In this case, protein H has been used to purify rabbit IgG, but the principle should be applicable to other M protein-ligand pairs.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Osmark P,Cedervall T,Pieters K,Akerström Bdoi
10.1016/s1046-5928(03)00130-xkeywords:
subject
Has Abstractpub_date
2003-08-01 00:00:00pages
301-3issue
2eissn
1046-5928issn
1096-0279pii
S104659280300130Xjournal_volume
30pub_type
杂志文章abstract：:Protein production can be improved if methods for soluble protein expression are developed. Interferon consensus (IFN-con) is used to treat hepatitis C. IFN-con has superior activity compared to other clinically used interferon α subtypes. However IFN-con is a challenging protein to produce in a soluble form using an ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2014.03.009
更新日期：2014-07-01 00:00:00
abstract：:Hereditary enzymopenic methemoglobinemia is a rare disease that predominantly results from defects in either the erythrocytic (type I) or microsomal (type II) forms of the enzyme NADH:cytochrome b5 reductase (EC 1.6.2.2). All 25 currently identified type I and type II methemoglobinemia mutants have been expressed in E...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00046-9
更新日期：2003-07-01 00:00:00
abstract：:A mammalian expression vector with features optimized for simple expression and purification of secreted proteins has been developed. This vector was constructed to facilitate X-ray crystallographic studies of cysteine-rich glycoproteins that are difficult to express by other means. Proteins expressed with this vector...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1331
更新日期：2000-12-01 00:00:00
abstract：:The gene encoding S. cerevisiae Kex2 protease derivative Kex2-667 (encoding the N-terminal 20th to 667th amino acid residues of Kex2 protease, containing the propeptide, catalytic domain, P domain and Ser/Thr enrichment region) and its 225th amino acid residue mutant K225L were overexpressed in Pichia pastoris. Protea...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105725
更新日期：2020-12-01 00:00:00
abstract：:κ-Carrageenase (EC3.2.1.83) is a class of glycoside hydrolase, which can be used for hydrolysis of κ-carrageenan to κ-carrageenan oligosaccharides. In this study, a bacterium, identified as Pseudoalteromonas sp. ZDY3 isolated from rotten algae, was capable to degrade κ-carrageenan. The κ-carrageenase produced by Pseud...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2020.105768
更新日期：2021-02-01 00:00:00
abstract：:Refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. We will review key parameters associated with (1) conformation of the protein solubilized from inc...
journal_title：Protein expression and purification
pub_type： 杂志文章,评审
doi：10.1016/s1046-5928(02)00641-1
更新日期：2003-03-01 00:00:00
abstract：:Structural biology places a high demand on proteins both in terms of quality and quantity. Although many protein expression and purification systems have been developed, an efficient and simple system which can be easily adapted is desirable. Here, we report a new system which combines improved expression, solubility ...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.09.008
更新日期：2009-02-01 00:00:00
abstract：:A simple and efficient method for hyperexpression in Escherichia coli and purification of capsid protein, p24, of human immunodeficiency virus type 1 (HIV-1) of both B- and C-subtypes is described. DNA-encoding p24 of C-subtype was cloned from C-subtype gag sequence which was obtained by PCR amplification using DNA ex...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1266
更新日期：2000-08-01 00:00:00
abstract：:Previously, the lysozyme gene of the Klebsiella phage K11 was partially sequenced in our lab. Using the sequence information the lysozyme gene of the Klebsiella phage K11 was amplified and cloned using the polymerase chain reaction of the pfu DNA polymerase. The nucleotide sequence of phage K11 lysozyme gene was deter...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2005.03.026
更新日期：2005-07-01 00:00:00
abstract：:α-N-Acetylgalactosaminidase (α-GalNAc-ase; EC.3.2.1.49) is an exoglycosidase specific for the hydrolysis of terminal α-linked N-acetylgalactosamine in various sugar chains. The cDNA corresponding to the α-GalNAc-ase gene was cloned from Aspergillus niger, sequenced, and expressed in the yeast Saccharomyces cerevisiae....
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2011.09.009
更新日期：2012-01-01 00:00:00
abstract：:Pokeweed antiviral protein (PAP)-I from the spring leaves of Phytolacca americana is a naturally occurring RNA-depurinating enzyme with broad-spectrum antiviral activity. Interest in PAP is growing due to its use as a potential anti-HIV agent. However, the clinical use of native PAP is limited due to inherent difficul...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1999.1181
更新日期：2000-03-01 00:00:00
abstract：:The human iron-binding protein lactoferrin (hLf) has been implicated in a number of important physiological pathways, including those regulating immune function and tumor growth. In an effort to develop an efficient system for production of recombinant hLf (rhLf) that is structurally and functionally equivalent to the...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2009.01.003
更新日期：2009-05-01 00:00:00
abstract：:A strategy for the purification and cleavage of chimeric recombinant proteins based on a genetically engineered metal-binding peptide and a human renin cleavage site is described. Vectors were constructed to direct the synthesis of chimeric human immunodeficiency virus (HIV) reverse transcriptase (RT) or beta-galactos...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/1046-5928(91)90073-r
更新日期：1991-04-01 00:00:00
abstract：:Alginate lyase digestion is an efficient way to degrade alginate into oligosaccharides, which are useful in various areas including chemistry, pharmacy and food industry. Here we determined the sequence of Vibrio sp. QY102 sourced alginate lyase, and set up its heterologous expression in E. coli. We improved its secre...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.09.002
更新日期：2019-01-01 00:00:00
abstract：:Proteins are essential throughout the biological and biomedical sciences and the purification strategies of proteins of interest have advanced over centuries. Elastin-like polypeptides (ELPs) are compound polymers that have recently been highlighted for their sharp and reversible phase transition property when heated ...
journal_title：Protein expression and purification
pub_type： 杂志文章,评审
doi：10.1016/j.pep.2018.09.006
更新日期：2019-01-01 00:00:00
abstract：:Hydrophobic charge-induction chromatography (HCIC) using 4-mercaptoethylpyridine (4-MEP) as the ligand is used to purify antibodies. The 4-MEP resin ligand has high affinity for antibodies, which makes it difficult to optimize the elution conditions. Recent studies showed that arginine is effective at eluting and puri...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2016.09.004
更新日期：2017-01-01 00:00:00
abstract：:Pigment epithelium-derived factor (PEDF) is a neurotrophic protein and a member of the serine protease inhibitor superfamily. Here we describe the identification of PEDF in bovine eyes and optimization of its purification from this natural source. We have developed a polyclonal antibody to recombinant human PEDF, Ab-r...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.1995.1060
更新日期：1995-08-01 00:00:00
abstract：:Rab GTPases are post-translationally geranylgeranylated on their C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase) and this modification is essential for their biological activity. Rab Escort Protein (REP) is a molecular chaperone that assists in the prenylation reaction carried out by RabGGTase. Muta...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00506-5
更新日期：2002-10-01 00:00:00
abstract：:Production of VCSM13 phage displaying a high density of CD147 ectodomain (CD147Ex) was achieved when culturing conditions were modulated. A phagemid expressing CD147Ex was constructed and used to produce phage display CD147Ex gpVIII fusion protein in TG1 Escherichia coli. Displaying of CD147Ex via gpVIII was successfu...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.08.019
更新日期：2003-12-01 00:00:00
abstract：:The recombinant human malonyl-CoA decarboxylase (hMCD) was overexpressed in Escherichia coli with and without the first 39 N-terminal amino acids via a cleavable MBP-fusion construct. Proteolytic digestion using genenase I to remove the MBP-fusion tag was optimized for both the full length and truncated hMCD. The apo-...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2003.11.023
更新日期：2004-04-01 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：:A method for purifying acylation stimulating protein (ASP) from porcine serum is described. The mRNA encoding ASP was cloned by reverse transcriptase-polymerase chain reaction which predicted a 76 residue peptide. Based on this sequence, we generated antisera to a C-terminal peptide (ASP(1-20)) which aided ASP purific...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(02)00019-0
更新日期：2002-07-01 00:00:00
abstract：:General procedures for the rapid, large-scale purification of recombinant Lactobacillus casei thymidylate synthase and its mutants have been established. An effective method employs sequential phosphocellulose and hydroxylapatite chromatography. Crude cell extracts are directly applied to phosphocellulose, and the enz...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(05)80039-7
更新日期：1992-10-01 00:00:00
abstract：:Receptor activator of NF-kappaB ligand (RANKL) is a type II transmembrane protein found on osteoblasts which functions as a major determinant of osteoclast differentiation and activation. RANKL mediates bone homeostasis through binding to the cognate ligand on osteoclasts, RANK, and a soluble decoy receptor, osteoprot...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1278
更新日期：2000-10-01 00:00:00
abstract：:S-locus protein kinase (SRK) is a receptor kinase that plays a critical role in self-recognition in the Brassicaceae self-incompatibility (SI) response. SRK is activated by binding of its ligand S-locus protein 11 (SP11) and subsequently induced phosphorylation of the intracellular kinase domain. However, a detailed a...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2015.09.020
更新日期：2017-03-01 00:00:00
abstract：:The promyelocytic leukemia (PML) gene is involved in the 15/17 chromosomal translocation of acute promyelocytic leukemia (APL). It encodes a nuclear phosphoprotein containing an alpha-helical coiled-coil domain with four heptad repeats. The heptad repeats consist of four clusters of hydrophobic amino acids that mediat...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/s1046-5928(03)00004-4
更新日期：2003-05-01 00:00:00
abstract：:We have developed a new expression vector, pcI(ts) ind(+), based upon the powerful rightward promoter of bacteriophage lambda, which is controlled by a temperature-sensitive and chemically-inducible version of the lambda repressor on the same plasmid. Locating the repressor gene on the plasmid makes this vector "porta...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2008.09.018
更新日期：2009-02-01 00:00:00
abstract：:Pullulanases are well-known starch-debranching enzymes that are widely used for hydrolysis of a-1,6-glycosidic linkages in starch, pullulan, amylopectin, and other oligosaccharides. Escherichia coli is a popular heterologous expression host for generating target enzymes. However, cells have to be disrupted to obtain t...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2018.09.011
更新日期：2019-03-01 00:00:00
abstract：:Cyclohexanone monooxygenase (CMO) is a soluble flavoenzyme originally isolated from Acinetobacter spp. which carries out Baeyer-Villiger reactions with cyclic ketone substrates. In the present study we cloned the Acinetobacter CMO gene and modified it for facile purification from heterologous expression systems by inc...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1006/prep.2000.1340
更新日期：2001-02-01 00:00:00
abstract：:High-level constitutive expression of xylanase GH11 from Penicillium occitanis Pol6 termed PoXyn2 was achieved using the methylotrophic yeast Pichia pastoris. The PoXyn2 cDNA encoding for a mature xylanase of 320 amino acids was subcloned into the pGAPZαA vector, to construct recombinant xylanse with six histidine res...
journal_title：Protein expression and purification
pub_type： 杂志文章
doi：10.1016/j.pep.2012.02.012
更新日期：2012-05-01 00:00:00
