Abstract:
:The human placenta is a rich raw material for production of many biopharmaceutical products. Here we describe a co-purification process for the production of four different proteins from haemolysed human placenta blood: IgG, catalase (Cat), superoxide dismutase (Sod) and albumin (Alb). The process can be divided in two parts: the common steps and the specific separation techniques for each protein. The common steps are: extraction, haemoglobin precipitation, concentration/diafiltration and the first Q-Sepharose chromatography step. At this chromatography step the process is branched: while IgG and Cat were recovered in the flow-through, Sod and Alb were eluted separately. IgG and Cat were separated in a second Q-Sepharose chromatography step during which IgG was recovered in the flow-through, whereas Cat bound to the resin. IgG was purified by S-Sepharose chromatography, followed by selective precipitation with n-octanoic acid, yielding about 0.4 g of IgG per kg of placenta. Cat was eluted at the second Q-Sepharose chromatography step and was purified by Blue Sepharose chromatography. A total of 1.8 x 10(6) units of Cat were recovered/kg of placenta, with a specific activity of 45000 units/mg of protein. Sod was further purified by S-Sepharose and Phenyl-Sepharose chromatography steps and recovered in the non-adsorbed fractions. The yield of Sod was 2.1 x 10(5) units/kg of placenta, with a specific activity of 1194 units/mg of protein. Alb purification was followed by a combined process including thermocoagulation and treatment with activated charcoal. The final step was Phenyl-Sepharose chromatography. The process yielded 3.1 g of Alb/kg of placenta. The described methodology was designed to be easily scaled-up for industrial production.
journal_name
Biotechnol Appl Biochemjournal_title
Biotechnology and applied biochemistryauthors
Grellet S,Martins EA,Maimoni Gonçalves V,Yague Lopes AP,Raw I,Cabrera-Crespo Jdoi
10.1042/ba20010023subject
Has Abstractpub_date
2001-12-01 00:00:00pages
135-42issue
3eissn
0885-4513issn
1470-8744journal_volume
34pub_type
杂志文章abstract::Lys and Leu were generally considered as the key amino acids for brewer's yeast during beer brewing. In the present study, peptide Lys-Leu and a free amino acid (FAA) mixture of Lys and Leu (Lys + Leu) were supplemented in 24 °P wort to examine their effects on physiological activity and fermentation performance of br...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1634
更新日期:2018-07-01 00:00:00
abstract::Mustard tuber wastewater is characterized by high salinity and high organic content that is potentially detrimental to the biological treatment system and affects the treatment efficiency accordingly. The experiment used the integrated bioreactor to reduce much of the organics in mustard tuber wastewater, and found th...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1396
更新日期:2016-07-01 00:00:00
abstract::The survival of inoculated microbes is critical for successful bioaugmentation in wastewater treatment. The influence of readily available nutrients (RANs) on the colonization of two functional bacteria, Pseudomonas putida M9, a strong biofilm-forming strain, and Comamonas testosteroni A3, a 3,5-dinitrobenzoic acid (3...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1314
更新日期:2015-09-01 00:00:00
abstract::Ethylene glycol bis (beta-aminoethyl ether)-N,N'-tetraacetic acid (EGTA)-calcium buffer is widely used in various calcium-dependent reactions where free calcium concentrations of 1 microM or less are desirable. The free calcium concentration is calculated from the association constant of EGTA . Ca2- and serves as the ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1986-10-01 00:00:00
abstract::Cardiolipin, the primary lipid hapten in the antigen suspension used for the detection of antitreponemal antibodies in the sera of syphilitic patients, was successfully coupled to glucose oxidase, peroxidase, and some other enzymes using different crosslinking agents. These complexes were used to replace the pure unco...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1986-12-01 00:00:00
abstract::Mutants with overexpression of α-acetolactate synthase (ALS), α-acetolactate decarboxylase, and acetoin reductase (AR), either individually or in combination, were constructed to improve 2,3-butanediol (2,3-BD) production in Klebsiella pneumoniae. The recombinant strains were characterized in terms of the enzyme activ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1217
更新日期:2014-11-01 00:00:00
abstract::The plant polymer lignin is the greatest source of aromatic chemical structures on earth. Hence, the chemically diverse lignin monomers are valuable raw materials for fine chemicals, materials synthesis, and food and flavor industries. However, extensive use of this natural resource is hampered by the large number of ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1222
更新日期:2014-11-01 00:00:00
abstract::The Staphylococcus aureus protein A (SpA) can be obtained through the culture of wild-type S. aureus and also as a recombinant protein in safe bacterial hosts. Several methods have been used to purify SpA among which ion-exchange chromatography, affinity chromatography, gel filtration, and per aqueous liquid chromatog...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章,评审
doi:10.1002/bab.1742
更新日期:2019-05-01 00:00:00
abstract::The purpose of the present study was to investigate the effects of garlic (Allium sativum L.) on the diabetic nephropathy and oxidative stress induced by STZ (streptozotocin) in rats. Diabetes was induced in Male Sprague-Dawley rats by administering a single intraperitoneal injection of STZ (60 mg/kg of body weight). ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/BA20080086
更新日期:2009-03-01 00:00:00
abstract::hES (human embryonic stem) cells hold tremendous potential in the newly emerging field of regenerative medicine, in addition to being a useful tool in basic scientific research and for pharmacological and cytotoxicity screening. However, an essential prerequisite for the future widespread application of hES cells are ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章,评审
doi:10.1042/BA20040161
更新日期:2005-04-01 00:00:00
abstract::Enzymatic synthesis of biodiesel showed advantageous characteristics in relation to other technologies once it works under bland conditions, no generation of wastewater, no occurrence of saponifications reactions and production of a biodiesel with high quality. Although many researches still apply immobilized lipases,...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章,评审
doi:10.1002/bab.1835
更新日期:2020-07-01 00:00:00
abstract::This review article focuses on the use of the methylotrophic yeast Pichia pastoris as a recombinant protein-expression system. P. pastoris is a useful system for the expression of milligram-to-gram quantities of a protein, which can be scaled up to fermentation to meet greater demands. Compared with mammalian cells, P...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章,评审
doi:
更新日期:1999-10-01 00:00:00
abstract::A rapid, simple and sensitive fluorometric assay method for the determination of nitrilase activity is described. 3-Cyanopyridine was hydrolysed to nicotinic acid by Rhodococcus rhodochrous and the liberated NH(3) was allowed to react with buffered o -phthaldialdehyde-2-mercaptoethanol solution (pH 7.4) to form a fluo...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/BA20020106
更新日期:2003-06-01 00:00:00
abstract::Single base mismatched oligonucleotides related to the rpoB gene of Mycobacterium tuberculosis, the mutations of which cause drug resistance of the infectious agent, were detected and discriminated using a surface plasmon resonance biosensor system. Thiol-modified oligonucleotides of the selected sequence (the probe) ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1101
更新日期:2013-07-01 00:00:00
abstract::Glutaraldehyde crosslinked, immunoprecipitated gulonolactone oxidase, injected intraperitoneally, has significant catalytic activity and is capable of providing long-term therapeutic benefit for the enzyme deficiency disease scurvy. The enzyme is tolerated even in repetitive doses. In the present study, however, we ha...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1986-04-01 00:00:00
abstract::Green fluorescent protein (GFP) is a globular protein used as biosensor and biomarker in medical and industrial fields. However, due to the expensive production costs of expressing proteins using high-cost inducers like isopropyl-β-d-1-thiogalactopyranoside (IPTG), the number of GFP applications are still scarce. This...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1749
更新日期:2019-07-01 00:00:00
abstract::In the present study, recombinant HCV (hepatitis C virus) core proteins enhanced the immune response elicited by a co-delivered DNA vaccine encoding HCV core and envelope proteins. A mixture of the plasmid pIDKE2 and Co.173, a protein comprising the first 173 amino acids of HCV core, in particular induces a strong hum...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/BA20050202
更新日期:2006-04-01 00:00:00
abstract::The pH optimum, temperature-dependence, thermal stability, substrate specificity and subsite affinities of the 66 kDa, pI 4.0 glucoamylase of the filamentous fungus Trichoderma reesei were determined. It had a pH optimum of 5.5 and a temperature optimum (5 min reaction time) of 70 degrees C with soluble starch as subs...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1111/j.1470-8744.1995.tb00333.x
更新日期:1995-04-01 00:00:00
abstract::Sephadex was derivatized consecutively with Staphylococcus Protein A (SpA) and cell-specific antibodies, and the binding of cells to the resulting material was examined. For comparison, cell binding to commercially obtained SpA-Sepharose was determined. Sephadex G-10, carboxylated by reaction with glycine and activate...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/ba20010062
更新日期:2002-02-01 00:00:00
abstract::Parkinson's disease (PD) is the most common neurodegenerative movement disorder and is characterized by the loss of dopaminergic neurons and the formation of eosinophilic intracytoplasmic inclusion bodies known as Lewy bodies. Although alpha-synuclein is known to be a pivotal factor implicated in the pathogenesis of P...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/ba20020004
更新日期:2002-08-01 00:00:00
abstract::Arylacetonitrile-hydrolyzing nitrilase (E.C. 3.5.5.5) of Alcaligenes sp. MTCC 10675 has been purified by up to 46-fold to homogeneity and 32% yield using ammonium sulfate fractionation, Sephacryl S-300 gel permeation, and anion exchange chromatography. The molecular weight of the native enzyme was estimated to be 520 ...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1192
更新日期:2014-07-01 00:00:00
abstract::Carbamate kinase has been prepared from Lactobacillus buchneri NCDO110. An approximately 91-fold increase in the specific activity of the enzyme was achieved. The purified extract exhibited a single band following polyacrylamide gel electrophoresis. The apparent molecular weight as determined by gel electrophoresis wa...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1986-02-01 00:00:00
abstract::Monoclonal antibodies specific to human prolactin were generated by an improved hybridoma technique. Following immunization with hormone conjugated with hemocyanin and cell fusion, hybrid cells were first cultured in a semisolid medium containing methylcellulose and later transferred to liquid medium for subculture. O...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1987-02-01 00:00:00
abstract::A three-layered simulation methodology is described that rapidly evaluates biomanufacturing process options. In each layer, inferior options are screened out, while more promising candidates are evaluated further in the subsequent, more refined layer, which uses more rigorous models that require more data from time-co...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1042/ba20070018
更新日期:2007-10-01 00:00:00
abstract::Heparin is a widely used clinical anticoagulant that is prepared from pig intestine. A contamination of heparin in 2008 has led to a reexamination of animal-derived pharmaceuticals. A bioengineered heparin prepared by bacterial fermentation and chemical and enzymatic processing is currently under development. This stu...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1042
更新日期:2012-11-01 00:00:00
abstract::Glycoprotein D (gD2) is the most important candidate antigen for herpes simplex virus type 2 (HSV-2) vaccine development. Establishment of a stable eukaryotic cell line to overexpress gD2 and an efficient purification process to purify is essential for the development of subunit vaccine against HSV-2. The DNA sequence...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1386
更新日期:2016-05-01 00:00:00
abstract::Chronic myelogenous leukemia (CML) is a hematologic malignancy characterized by excessive growth of myeloid cells and their progenitors. The proportion of spectrin dimers compared to tetramers extracted from membranes at 4 degrees C, under low ionic strength conditions, increased in CML erythrocytes. These also displa...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章,评审
doi:
更新日期:1990-10-01 00:00:00
abstract::Expression of the VP2 gene of autonomous parvoviruses in insect cells with the use of the baculovirus system has led to the production of virus-like particles (VLPs) formed by the self-assembly of VP2. These VLPs are expressed at high levels and can easily be purified by salt fractionation. They are highly immunogenic...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1999-04-01 00:00:00
abstract::A novel iron superoxide dismutase (Fe-SOD) gene from Jatropha curcas was cloned and expressed in Escherichia coli BL21 (DE3). This recombinant enzyme was isolated by a combined procedure involving immobilized metal-ion affinity chromatography and ion-exchange chromatography. The apparent molecular mass of this purifie...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:10.1002/bab.1030
更新日期:2012-09-01 00:00:00
abstract::alpha-Cyclodextrin (CD) was converted by Bacillus macerans cyclodextrin glucanotransferase into highly insoluble high-amylose starch with a low average degree of polymerization (56-73), in yields as high as 78% over a wide range of temperatures (25-70 degreesC). Ability to undergo this conversion was highly concentrat...
journal_title:Biotechnology and applied biochemistry
pub_type: 杂志文章
doi:
更新日期:1998-12-01 00:00:00