Abstract：
:Deficiency of the lysosomal glucocerebrosidase (GCR) enzyme results in Gaucher's disease, the most common inherited storage disorder. Treatment consists of enzyme replacement therapy by the administration of recombinant GCR produced in Chinese hamster ovary cells. The production of anti-GCR antibodies has already been described with placenta-derived human GCR that requires successive chromatographic procedures. Here, we report a practical and efficient method to obtain anti-GCR polyclonal antibodies against recombinant GCR produced in Escherichia coli and further purified by a single step through nickel affinity chromatography. The purified GCR was used to immunize BALB/c mice and the induction of anti-GCR antibodies was evaluated by enzyme-linked immunosorbent assay. The specificity of the antiserum was also evaluated by western blot analysis against recombinant GCR produced by COS-7 cells or against endogenous GCR of human cell lines. GCR was strongly recognized by the produced antibodies, either as cell-associated or as secreted forms. The detected molecular masses of 59-66 kDa are in accordance to the expected size for glycosylated GCR. The GCR produced in E. coli would facilitate the production of polyclonal (shown here) and monoclonal antibodies and their use in the characterization of new biosimilar recombinant GCRs coming in the near future.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Novo JB,Oliveira ML,Magalhães GS,Morganti L,Raw I,Ho PLdoi
10.1007/s12033-010-9303-4subject
Has Abstractpub_date
2010-11-01 00:00:00pages
279-86issue
3eissn
1073-6085issn
1559-0305journal_volume
46pub_type
杂志文章abstract：:This article reviews some of the issues involved in evaluating vaccines in humans. Vaccine trials are required for licensure and are essential for demonstrating a vaccine's safety and protective efficacy. The formal framework of phase I, II, and III trials is described, with particular emphasis on the choice of hypoth...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:17:1:43
更新日期：2001-01-01 00:00:00
abstract：:Pyrococcus woesei (DSM 3773) beta-galactosidase gene amplified by polymerase chain reaction was cloned into KpnI and HindIII binding sites of pET-30LIC expression plasmid. The obtained pGal2 (6785 bp) transcription vector was then transferred to Escherichia coli B121 (DE3) cells. High identity (99.9%) of DNA sequences...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02740841
更新日期：1998-12-01 00:00:00
abstract：:Adenoviruses are attracting increasing attention as general purpose mammalian cell expression vectors, as recombinant vaccines, and potentially as vectors for gene therapy. Not only is the adenovirus genome relatively easy to manipulate by recombinant DNA techniques, but adenovirus vectors are relatively stable, grow ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789331
更新日期：1995-06-01 00:00:00
abstract：:A shuttle vector pZL1 which can replicate both in Gluconobacter oxydans and Escherichia coli was constructed based on G. oxydans DSM2003 cryptic plasmid pGOX3, a homology of G. oxydans 621H pGOX3, and E. coli cloning vector pUC18. It was found to be stably maintained in G. oxydans during the serial subcultures in the ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-010-9293-2
更新日期：2010-11-01 00:00:00
abstract：:This article presents a simple and rapid method for removal of unincorporated label and proteins from DNA sequencing reactions by using Wizard purification resin. This method can be successfully applied for preparation of end-labeled oligonucleotides free of unincorporated label, which is important in experiments (inc...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789066
更新日期：1996-04-01 00:00:00
abstract：:Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing the inhibiting transgene product. A gene regulation system based on t...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02686012
更新日期：2007-03-01 00:00:00
abstract：:A bacterial artificial chromosome library of the causal agent of the Black Sigatoka leaf spot disease of banana and plantain, Mycosphaerella fijiensis, has been constructed using a non-sphaeroplasting technique and characterized using both homologous and heterologous probes. After first and a second size selection of ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-0016-2
更新日期：2007-05-01 00:00:00
abstract：:Considering the fact that Prays oleae is one of the most pathogenic insects to the olive tree in the Mediterranean basin, particularly in Tunisia, the mode of action of Cry insecticidal toxins of Bacillus thuringiensis kurstaki in Prays oleae midgut was investigated. The proteolysis of Bacillus thuringiensis delta-end...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02686109
更新日期：2007-02-01 00:00:00
abstract：:In this study, we investigated expression and dimerization of an ER-associated degradation (ERAD) substrate, a null Hong Kong variant of α-1-antitrypsin (NHK) using immunoblotting assay and a novel NanoLuc complementary reporter system called the NanoBiT (NB) assay. This NB-tagged NHK made it possible to monitor the i...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-018-0092-5
更新日期：2018-08-01 00:00:00
abstract：:The implementation of efficient technologies for the production of recombinant mammalian membrane receptors is an outstanding challenge in understanding receptor-ligand actions and the development of therapeutic antibodies. In order to improve the solubility of recombinant extracellular domains of human membrane recep...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:33:3:199
更新日期：2006-07-01 00:00:00
abstract：:High-quality RNA preparations are critical for further applications such as reverse transcriptase-polymerase chain reaction (RT-PCR) transcript amplifications, and elaboration of cDNA and expressed sequence tag libraries. Melanins are phenolic compounds present in many fungi and apparently play key roles in fungi path...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:34:1:45
更新日期：2006-09-01 00:00:00
abstract：:The major online single nucleotide polymorphism (SNP) databases freely available as research tools for genetic analysis are explained, reviewed, and compared. An outline is given of the search strategies that can be used with the most extensive current SNP databases: National Centre for Biotechnology Information (NCBI...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/mb:35:1:65
更新日期：2007-01-01 00:00:00
abstract：:A short (43-bp) A/T-rich stretch of DNA located in the intergenic region between the baiA2 and baiF genes from Eubacterium sp. strain VPI 12708 was amplified by polymerase chain reaction (PCR) and inserted in front of the Shine-Dalgarno (SD) sequences of three inefficiently-expressed Eubacterium sp. strain VPI 12708 g...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02789174
更新日期：1999-02-01 00:00:00
abstract：:IgG class antibodies express catalytic activities rarely and at very low levels. Here, we studied polyclonal IgA and IgG preparations from healthy human sera and saliva for the ability to hydrolyze model peptidyl-aminomethylcoumarin (peptide-AMC) substrates. These substrates permit objective evaluation of the catalyti...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-0003-7
更新日期：2007-06-01 00:00:00
abstract：:A synthetic gene encoding a single chain Fv fragment of an antibody directed against the nuclear inclusion a (NIa) protein of potato virus Y (PVY) was used to transform two commercial potato cultivars (Claustar and BF15). The NIa protease forms the nuclear inclusion body A and acts as the major protease in the cleavag...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:33:2:133
更新日期：2006-06-01 00:00:00
abstract：:The diversity and complexity of bioinformatics tools currently available for protein sequence analysis can make it difficult to know where to begin when presented with a new sequence. In this article, we present a protocol outlining one approach to sequence analysis that should give as comprehensive a picture as possi...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-007-0032-2
更新日期：2007-07-01 00:00:00
abstract：:The two asparagine-linked glycosylation sites of recombinant coagulation factor VIIa have been characterized by glycosidase digestions, size-exclusion chromatography (SEC), and mass spectrometry (MS). Nine structures were characterized as core fucosylated bi-and triantennary structures with 0-3 sialic-acid residues wh...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：
更新日期：1998-06-01 00:00:00
abstract：:Optimization of gene coding-sequence, including preferred codon usage and removal of cryptic splice sites and mRNA-destabilizing motifs, has been shown to improve recombinant protein production of different proteins. Here, we present data to show that gene optimization can also be used to improve the production of a c...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:34:2:151
更新日期：2006-10-01 00:00:00
abstract：:In this study, host-specific forms of the blast pathogen Magnaporthe oryzae in sub-Saharan Africa (SSA) were characterised from distinct cropping locations using a combination of molecular and biological assays. Finger millet blast populations in East Africa revealed a continuous genetic variation pattern and lack of ...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-011-9429-z
更新日期：2012-02-01 00:00:00
abstract：:HOX genes are highly conserved transcription factors that determine the identity of cells and tissues along the anterior-posterior body axis in developing embryos. Aberrations in HOX gene expression have been shown in various tumors. However, the correlation of HOX gene expression patterns with tumorigenesis and cance...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-013-9682-4
更新日期：2014-01-01 00:00:00
abstract：:The biosensor field has grown enormously since the first demonstration of the biosensor concept by Leland C. Clark, Jr. in 1962. Today's biosensor market is dominated by glucose biosensors, mass-produced enzyme electrodes for the rapid self-diagnosis of blood glucose levels by diabetes sufferers. Here we take a histor...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:32:3:249
更新日期：2006-03-01 00:00:00
abstract：:De novo design provides an in silico toolkit for the design of novel small molecular structures to a set of specified structural constraints. With the avalanche of bioinformatics data, de novo design is ideally suited for exploring molecules that could be useful for chemical genomics. The design process involves manip...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-007-0037-x
更新日期：2007-11-01 00:00:00
abstract：:Identification of expressed sequence tags (ESTs) in large genomic segments is an important step in positional cloning and genomic mapping studies. A simple and efficient polymerase chain reaction (PCR)-based approach is described here to identify coding sequences in large genomic fragments of DNA cloned in vectors suc...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/BF02760779
更新日期：1997-12-01 00:00:00
abstract：:Murraya koenigii (L.) Spreng. (Rutaceae), is an aromatic plant and much valued for its flavor, nutritive and medicinal properties. In this study, three DNA fingerprinting methods viz., random amplification of polymorphic DNA (RAPD), directed amplification of minisatellite DNA (DAMD), and inter-simple sequence repeat (...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9500-4
更新日期：2013-02-01 00:00:00
abstract：:Epoxide hydrolases (EHs; 3.3.2.x) catalyze the enantioselective ring opening of racemic epoxides to the corresponding enantiopure vicinal diols and remaining equivalent unreacted epoxides. These epoxides and diols are used for the synthesis of chiral drug intermediates. With an upsurge in the methods for identificatio...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-017-9996-8
更新日期：2017-03-01 00:00:00
abstract：:Monoclonal antibodies (Mabs) against human chorionic gonadotropin hormone (hCG) were raised by hybridoma technology using Sp2/0 myeloma cells as fusion partner. Sixty-five percent of the total culture wells exhibited hybrid growth and 8% of the total wells (13 culture wells) contained anti-hCG secreting hybrids. A pos...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1385/MB:17:2:119
更新日期：2001-02-01 00:00:00
abstract：:Despite the known importance of the composition of culture media and culture conditions on Bacillus thuringiensis growth and toxicity, very few reviews are concerned with this subject. This article reviews some aspects of the microbiology of Bacillus thuringiensis, and how toxicity is affected by the composition of gr...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/BF02907471
更新日期：1995-08-01 00:00:00
abstract：:Monoclonal antibodies (mAbs) are the fastest growing class of biopharmaceuticals reflecting their diverse applications in research and the clinic. The correct glycosylation of mAbs is required to elicit effector functions such as complement-dependent and antibody-dependent cell-mediated cytotoxicity, although these ma...
journal_title：Molecular biotechnology
pub_type： 杂志文章
doi：10.1007/s12033-012-9531-x
更新日期：2013-03-01 00:00:00
abstract：:Ribosome display is a powerful engineering research tool for the high-throughput selection of peptides or proteins, which results in the generation of high-performance binders against nearly any antigen of interest. As a cell-free display system, ribosome display has been well developed with many outstanding achieveme...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1007/s12033-018-0133-0
更新日期：2019-01-01 00:00:00
abstract：:Technological achievements in the last 5 to 10 yr and their application to sequencing and polymorphism discovery in the human genome have fostered a renewed interest in the genetic basis of drug response. Consequently, the field of pharmacogenetics/pharmacogenomics has been gaining momentum, fueled not only on technol...
journal_title：Molecular biotechnology
pub_type： 杂志文章,评审
doi：10.1385/MB:25:3:275
更新日期：2003-11-01 00:00:00
