Application of polyacrylamide slab gel electrophoresis to the analysis and small-scale purification of amyloid proteins

1998 ◽  
Vol 372 (1-2) ◽  
pp. 161-172 ◽  
Author(s):  
B. Kaplan
Keyword(s):  
Gel Electrophoresis ◽  
Small Scale ◽  
Amyloid Proteins

Optimized Large Scale Synthesis of Phosphorothioate Oligonucleotides on PS-PEG Supports

1997 ◽  
Vol 52 (1) ◽  
pp. 110-116
Author(s):  
Michael Gerster ◽  
Martin Maier ◽  
Nils Clausen ◽  
Jens Schewitz ◽  
Ernst Bayer
Keyword(s):  
Gel Electrophoresis ◽  
Continuous Flow ◽  
Large Scale ◽  
Electrospray Mass Spectrometry ◽  
Small Scale ◽  
Batch Mode ◽  
Crucial Step ◽  
Capillary Gel Electrophoresis ◽  
Phosphoramidite Chemistry ◽  
Large Scale Synthesis

Sulphurization is a crucial step during synthesis of phosphorothioate oligonucleotides. Insufficient reaction leads to inhomogeneous products with phosphodiester defects and subsequently to destabilization of the oligomers in biological media. To achieve a maximum extent of sulphur incorporation, various sulphurizing agents have been investigated. Solely, the use of Beaucage reagent provided satisfactory results on PS-PEG supports. Based on our investigations in small scale synthesis (1 μmol) with continuous-flow technique, upscaling to the 0.1-0.25 mmolar range has been achieved using a peptide synthesizer. The syntheses were performed in batch mode with standard phosphoramidite chemistry. Additionally, large scale synthesis of a phosphodiester oligonucleotide has been carried out on PS-PEG with optimized protocols and compared to small scale synthesis on different supports. Products were analysed by 31P NMR, capillary gel electrophoresis and electrospray mass spectrometry. An extent of sulphurization of 99% and coupling effiencies of more than 99% were obtained and the products proved to have similar purity compared to small scale syntheses on CPG

Characterization of Listeria monocytogenes Isolates from 50 Small-Scale Austrian Cheese Factories

2006 ◽  
Vol 69 (6) ◽  
pp. 1297-1303 ◽  
Author(s):  
M. WAGNER ◽  
F. ELISKASES-LECHNER ◽  
P. RIECK ◽  
I. HEIN ◽  
F. ALLERBERGER
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Wide Distribution ◽  
Pulsed Field Gel Electrophoresis ◽  
Annual Production ◽  
Personal Hygiene ◽  
Small Scale ◽  
Pulsed Field ◽  
High Prevalence

One hundred eighty-one small-scale cheese factories (annual production <100,000 kg) were tested for the presence of Listeria monocytogenes in cheese and smear samples from 1997 to 2000. In total, 2615 samples were drawn. Fifty (27.6%) of 181 enterprises yielded L. monocytogenes. From 14 of the cheese-making facilities, we obtained more than four L. monocytogenes isolates. A total of 182 mostly cheese- and smear-borne L. monocytogenes strains were characterized by serotyping and pulsed-field gel electrophoresis. In 12 of 14 cheese factories, over half of the L. monocytogenes isolates were genetically indistinguishable by pulsetype. On average, genetically indistinguishable isolates were recovered for 11.9 months. Regarding serotypes, 27.3% of the isolates were of serovar 4b. Inadequate personal hygiene could explain the high prevalence of serovar 4b isolates in small-scale cheesemaking facilities. Forty-two percent of the serovar 4b isolates recovered from epidemiologically unlinked facilities (in comparison to 40 and 29% of the 1/2a and 1/2b isolates, respectively) were genetically indistinguishable from at least one other isolate. Indistinguishable serovar 1/2a and 1/2b isolates belonged to five and six different pulsetypes, respectively, whereas serovar 4b isolates belonged to only two pulsetypes. This finding suggested a wide distribution of genetically homologous serovar 4b isolates among the facilities tested in our study.

scholarly journals Tracing of Listeria monocytogenes Contamination Routes in Fermented Sausage Production Chain by Pulsed-Field Gel Electrophoresis Typing

Foods ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 198 ◽  
Author(s):  
Valerij Pažin ◽  
Dean Jankuloski ◽  
Lidija Kozačinski ◽  
Vesna Dobranić ◽  
Bela Njari ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Gel Electrophoresis ◽  
Environmental Samples ◽  
Detection System ◽  
Pulsed Field Gel Electrophoresis ◽  
Small Scale ◽  
Group Method ◽  
Fermented Sausage ◽  
Production Chain ◽  
Pulsed Field

In this study, the presence of Listeria monocytogenes was assessed along the production process of fermented sausages in a small-scale facility. Following the isolation of the pathogen from the final product (ISO 11290-1), retrospective sampling was performed during the production of a new batch of sausages, including raw materials, casings, additives, sausage mixtures, sausages during fermentation, and environmental samples. L. monocytogenes was recovered from the following sampling points: the defrosting room and the cuttering, mixing, stuffing, and fermentation phases. Ten strains were isolated, molecularly confirmed as L. monocytogenes by means of a molecular detection system, and subjected to pulsed-field gel electrophoresis (PFGE) typing. On the basis of an unweighted pair group method with arithmetic mean (UPGMA) dendrogram from Ascl pulsotypes, the strains were indistinguishable (no band difference). The same pulsotypes of strains present in both batches of sausages, as well as in environmental samples, indicated the persistence of L. monocytogenes in the sausage production unit.

The purification of porcine haptoglobin by affinity chromatography with concanavalin A – Sepharose

1976 ◽  
Vol 54 (11) ◽  
pp. 999-1001 ◽  
Author(s):  
Frank A. Terpstra ◽  
David B. Smith
Keyword(s):  
Affinity Chromatography ◽  
Concanavalin A ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Gel Filtration ◽  
Polyacrylamide Gel ◽  
Small Scale ◽  
Scale Separation

Methods designed for the isolation of human haptoglobin (Hp) were found insufficient when applied to pig plasma due to the formation of a material tentatively identified as albumin dimer. Small scale separation is possible by preparative polyacrylamide gel electrophoresis. Larger scale purification from nonglycoprotein contaminants such as albumin dimer is achieved by affinity chromatography using immobilized concanavalin A. Porcine haptoglobin is microheterogeneous. More than 14 components, partially resolveable by gel filtration, were detected.

Purification and some properties of a thermostable DNA polymerase from a Thermotoga species

1990 ◽  
Vol 68 (11) ◽  
pp. 1292-1296 ◽  
Author(s):  
H. D. Simpson ◽  
T. Coolbear ◽  
M. Vermue ◽  
R. M. Daniel
Keyword(s):  
Dna Polymerase ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Thermostable Enzyme ◽  
Maximum Activity ◽  
Polyacrylamide Gel ◽  
Size Exclusion ◽  
Small Scale ◽  
Ph Optimum

A stable DNA polymerase (EC 2.7.7.7) has been purified from the extremely thermophilic eubacterium Thermotoga sp. strain FjSS3-B.1 by a five-step purification procedure. First, the crude extract was treated with polyethylenimine to precipitate nucleic acids. The endonuclease activity coprecipitated. DEAE-Sepharose, CM-Sephrarose, and hydroxylapatite column chromatography were used to purify the preparation. As a final step on a small scale, preparative sodium dodecyl sulfate (SDS) – polyacrylamide gel electrophoresis was used. The purified DNA polymerase exhibited a molecular weight of 85 000, as determined by both SDS–polyacrylamide gel electrophoresis and size-exclusion chromatography. Its pH optimum was in the range pH 7.5–8. When assayed over the temperature range 30–80 °C, the maximum activity in a 30-min assay was at 80 °C. The enzyme was moderately thermostable and exhibited half-lives of 3 min at 95 °C and 60 min at 50 °C in the absence of substrate. Several additives such as Triton X-100 enhanced thermostability. During storage at 4 °C and −70 °C, the stability of the enzyme was improved by the addition of gelatin.Key words: DNA polymerase, thermostable enzyme, Thermotoga.

Small-Scale Analysis of O-Linked Oligosaccharides from Glycoproteins and Mucins Separated by Gel Electrophoresis

2002 ◽  
Vol 74 (23) ◽  
pp. 6088-6097 ◽  
Author(s):  
Benjamin L. Schulz ◽  
Nicolle H. Packer ◽  
Niclas G. Karlsson
Keyword(s):  
Gel Electrophoresis ◽  
Small Scale ◽  
Scale Analysis
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