Cell-free protein synthesis: advances on production process for biopharmaceuticals and immunobiological products.

abstract：:A dot immunoblotting technique has been developed to estimate the relative expression levels of tagged recombinant human proteins in mammalian cell culture media. Variations in sample denaturation, blocking agents and membrane composition and treatment were used to optimize the signal-to-noise ratio of the defined pro...

journal_title：BioTechniques

authors： Ellsworth JL,Hamacher N,Bagwell N,West JW

更新日期：2000-05-01 00:00:00

abstract：:Planar lipid bilayers represent a versatile platform for studying the functions of various membrane proteins as well as the development of biosensors. Despite the continuing technological progress in the fabrication of low-noise bilayer setups with mechanically and electrically stable planar bilayers, there is still a...

journal_title：BioTechniques

authors： Novák P,Gaburjáková M,Zahradník I

更新日期：2007-03-01 00:00:00

abstract：:The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial...

journal_title：BioTechniques

authors： Yusuf M,Chen B,Hashimoto T,Estandarte AK,Thompson G,Robinson I

更新日期：2014-12-01 00:00:00

abstract：:A method for cloning single-stranded oligonucleotides in a plasmid vector has been developed. The method relies on ligation of the oligonucleotide into suitable restriction enzyme sites of the cloning vector such that the site at the 5' end has a 5' overhang [for example, a Bgl II site (A decreases GATCT)], and the si...

journal_title：BioTechniques

authors： Mounts P,Wu TC,Peden K

更新日期：1989-04-01 00:00:00

abstract：:We present a tool for dispensing very low volumes (20 nL or more) of ultra high viscosity (UHV) medical-grade alginate hydrogels. It uses a modified piezo-driven micrometering valve, integrated into a versatile system that allows fast prototyping of encapsulation procedures and scaffold production. Valves show excelle...

journal_title：BioTechniques

authors： Gepp MM,Ehrhart F,Shirley SG,Howitz S,Zimmermann H

更新日期：2009-01-01 00:00:00

abstract：:The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converte...

journal_title：BioTechniques

authors： Katz D,Ito E,Lau KS,Mocanu JD,Bastianutto C,Schimmer AD,Liu FF

更新日期：2008-02-01 00:00:00

abstract：:Recombinant retroviral vectors usually encode the genes of interest in place of the viral structural genes, which must be provided in trans. These viruses are therefore defective for replication: infected cells cannot produce progeny virus. However, in some cases it may be desirable to generate virus from an infected ...

journal_title：BioTechniques

authors： Corbley MJ,Cherington V,Feig LA,Cooper GM,Roberts TM

更新日期：1994-12-01 00:00:00

abstract：:The study of gene regulation in cardiac myocytes requires a reliable in vitro model. However, monolayer cultures used for this purpose are typically not exposed to electrical stimulation, though this has been shown to strongly affect cardiomyocyte gene expression. Based on pacemakers for clinical use, we developed an ...

journal_title：BioTechniques

authors： Martherus RS,Zeijlemaker VA,Ayoubi TA

更新日期：2010-01-01 00:00:00

abstract：:We have developed a rapid [3H]colchicine competition-binding scintillation proximity assay (SPA) to evaluate antimitotic compounds that bind to the colchicine-binding site on tubulin. The premise of our assay is that compounds will compete with radiolabeled colchicine for the tubulin-binding domain. Biotin-labeled tub...

journal_title：BioTechniques

authors： Tahir SK,Kovar P,Rosenberg SH,Ng SC

更新日期：2000-07-01 00:00:00

abstract：:The purpose of this research was to compare three bioreactor systems for the pilot-scale production of monoclonal antibodies (MAbs). We needed to produce gram quantities of murine MAbs against human prostatic acid phosphatase for use in fragmentation, radiolabeling and in vivo radio-imaging studies. The stable hybrido...

journal_title：BioTechniques

authors： Kurkela R,Fraune E,Vihko P

更新日期：1993-10-01 00:00:00

abstract：:A fluorescent in situ DNA hybridization assay employing a biotinylated DNA probe was used to visualize single copies of human immunodeficiency virus (HIV) proviral DNA in the nuclei and metaphase chromosomes of infected cells. In clonal cell lines that contain either one or two copies of proviral DNA, the efficiency o...

journal_title：BioTechniques

authors： Spadoro JP,Payne H,Lee Y,Rosenstraus MJ

更新日期：1990-08-01 00:00:00

abstract：:In the design of oligonucleotide sequences for targeting DNA or RNA sequences, it can be difficult to identify sequences that will hybridize only to the intended target. The term "sequence-specific" or "sequence-nonspecific" is often used to describe the interactions of an oligonucleotide with a mixture of DNA or RNA....

journal_title：BioTechniques

authors： Hyndman D,Cooper A,Pruzinsky S,Coad D,Mitsuhashi M

更新日期：1996-06-01 00:00:00

abstract：:When PCR is carried out in a polyacrylamide gel, each target molecule forms a molecular colony that comprises many copies of the original template. By counting the number of colonies, one can directly determine the target titer, with 100% of the DNA molecules and approximately 15% of the RNA molecules being detected. ...

journal_title：BioTechniques

authors： Chetverina HV,Samatov TR,Ugarov VI,Chetverin AB

更新日期：2002-07-01 00:00:00

abstract：:Francisella tularensis is one of the most deadly bacterial agents, yet most of the genetic determinants of pathogenesis are still unknown. We have developed an efficient targeted mutagenesis strategy in the model organism F. tularensis subsp. novicida by utilizing universal priming of optimized antibiotic resistance c...

journal_title：BioTechniques

authors： Liu J,Zogaj X,Barker JR,Klose KE

更新日期：2007-10-01 00:00:00

abstract：:The increase of information in biology makes it difficult for researchers in any field to keep current with the literature. The MEDLINE database of scientific abstracts can be quickly scanned using electronic mechanisms. Potentially interesting abstracts can be selected by matching words joined by Boolean operators. H...

journal_title：BioTechniques

authors： Perez-Iratxeta C,Keer HS,Bork P,Andrade MA

更新日期：2002-06-01 00:00:00

abstract：:Determining the primary sequences of informational macromolecules is no longer a limiting factor for our ability to completely understand the biological functioning of cells and organisms. Similarly, our understanding of transcriptional regulation (transcriptomics) has been greatly enhanced by the availability of micr...

journal_title：BioTechniques

authors： Kandpal R,Saviola B,Felton J

更新日期：2009-04-01 00:00:00

abstract：:Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene produ...

journal_title：BioTechniques

authors： Gutheil JC,Mane SM,Bass KA,Lee EJ,Needleman SW

更新日期：1990-08-01 00:00:00

abstract：:Directed evolution is poised to change small molecule discovery and provide greater access to "drug space." Sarah Webb looks into the evolving drug discovery landscape. ...

journal_title：BioTechniques

authors： Webb S Ph D

更新日期：2017-12-01 00:00:00

abstract：:A procedure for amplification by PCR of reproducible allele markers for amplified fragment length polymorphism (Amp-FLP) analysis is presented. We have prepared markers for the allelic products of the VNTR loci D1S80 (MCT118) and D17S30 (YNZ22) and for the hypervariable VNTR locus close to the 3' end of the apolipopro...

journal_title：BioTechniques

authors： Sajantila A,Puomilahti S,Johnsson V,Ehnholm C

更新日期：1992-01-01 00:00:00

abstract：:Gene expression analysis using high-density cDNA or oligonucleotide arrays is a rapidly emerging tool for transcriptomics, the analysis of the transcriptional state of a cell or organ. One of the limitations of current methodologies is the requirement of a relatively large amount of total or polyadenylated RNA as star...

journal_title：BioTechniques

authors： Scherer A,Krause A,Walker JR,Sutton SE,Serón D,Raulf F,Cooke MP

更新日期：2003-03-01 00:00:00

abstract：:We have developed a novel vector pTCS, as a tool for efficient selection of open reading frame (ORF)-containing inserts. In pTCS clones containing an insert with an ORF a downstream marker gene (immE3, conferring resistance to colicin) is activated via translational coupling with the insert, and transformed cells can ...

journal_title：BioTechniques

authors： Ohashi-Kunihiro S,Yohda M,Masaki H,Machida M

更新日期：2007-12-01 00:00:00

abstract：:A method was developed for the detection of bacterial mRNAs using reverse transcriptase followed by the polymerase chain reaction (PCR) and Southern blot analysis. The method involves brief inhibition of protein synthesis with chloramphenicol, followed by reverse transcription, PCR amplification of cDNA and Southern b...

journal_title：BioTechniques

authors： Mahbubani MH,Bej AK,Miller RD,Atlas RM,DiCesare JL,Haff LA

更新日期：1991-01-01 00:00:00

abstract：:A simple, nondestructive PCR-based screening method has been developed for identifying putative transgenic soft white winter wheat (Triticum aestivum L.) carrying the coat protein gene of wheat streak mosaic virus. Removal of the endosperm end of individual seed provided sufficient material for DNA extraction and PCR....

journal_title：BioTechniques

authors： McCarthy PL,Hansen JL,Zemetra RS,Berger PH

更新日期：2002-03-01 00:00:00

abstract：:The polymorphism of the fourth component of human serum complement (C4) is well established at the proteinic level; at the DNA level in the analysis of C4A and C4B gene polymorphism, the PCR technique is not widely and routinely used because it is time consuming and still presents reproducibility problems. This is a s...

journal_title：BioTechniques

authors： Zorzetto M,Campo I,Cortelazzo AG,Panelli S,Cuccia M

更新日期：2001-05-01 00:00:00

abstract：:Few studies have compared the quantification of mRNA by DNA microarray to the results obtained by reverse transcription PCR (RT-PCR). In this study, mRNA was collected from the healing femoral fracture callus of adult and juvenile rats at various times after fracture. Ten samples were measured by both methods for 26 g...

journal_title：BioTechniques

authors： Etienne W,Meyer MH,Peppers J,Meyer RA Jr

更新日期：2004-04-01 00:00:00

abstract：:Microarray expression analysis has become one of the most widely used functional genomics tools. Efficient application of this technique requires the development of robust and reproducible protocols. We have optimized all aspects of the process, including PCR amplification of target cDNA clones, microarray printing, p...

journal_title：BioTechniques

authors： Hegde P,Qi R,Abernathy K,Gay C,Dharap S,Gaspard R,Hughes JE,Snesrud E,Lee N,Quackenbush J

更新日期：2000-09-01 00:00:00

abstract：:We report a modification of a liquid hybridization method that serves as a rapid, sensitive alternative to Southern hybridization for the analysis of polymerase chain reaction (PCR) products. An aliquot of the completed PCR is mixed with an internally nested, end-labeled oligonucleotide probe, and one cycle of PCR is ...

journal_title：BioTechniques

authors： Parker JD,Burmer GC

更新日期：1991-01-01 00:00:00

abstract：:We have developed an automated purification method for dye-terminator-based DNA sequencing products using a magnetic bead approach. This 384-well protocol generates sequence fragments that are essentially free of template DNA, salt, and excess dye-terminator products. In comparison with traditional ethanol precipitati...

journal_title：BioTechniques

authors： Elkin C,Kapur H,Smith T,Humphries D,Pollard M,Hammon N,Hawkins T

更新日期：2002-06-01 00:00:00

abstract：:Data are presented illustrating the optimum concentration range of reverse transcription PCR-generated products under 500 bp for accurate base calling with direct automated DNA sequencing. A 357-bp fragment of the human estrogen receptor, which includes the DNA binding domain of the protein, was used as a representati...

journal_title：BioTechniques

authors： Hyder SM,Hu C,Needleman DS,Sonoda Y,Wang XY,Baker VV

更新日期：1994-09-01 00:00:00

abstract：:We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polyme...

journal_title：BioTechniques

authors： Gupta S,Arora K,Sampath A,Khurana S,Singh SS,Gupta A,Chaudhary VK

更新日期：1999-08-01 00:00:00