Everything you need to know about Ultrapure Water that Improves Results In 2D Gel Electrophoresis

Overview

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• Source: Sartorius


• Date: 04 Jan,2022

The U.S. Food and Drug Administration (FDA), launched an initiative called Pharmaceutical CGMMPs for 21st Century — Risk-Based Approach in August 2002. The FDA’s recommendations for the use of technological advances in pharmaceutical production, as well as the implementation of modern quality management and risk management tools and concepts were highlighted in this initiative. The FDA’s constant striving to achieve the goals set forth in this initiative led to the publication of several guidance that give industry better tools for modernization.

A recent FDA investigation into a multistate epidemic has revealed that the water-based bacteria burkholderia cepacia was found in more than 10 lots oral liquid docusate salt linked to an older water system at a Florida-based contract manufacturing organization (CMO).

This is a stark example of the seriousness of the situation in an industry where many manufacturers seem to prefer proven, older technologies to newer ones. Water’s incredible solvent properties allow it to mix with many substances and dissolve them.

This means that lab water sources can contain many contaminants. Even tiny amounts of impurities can cause problems in a number of scientific applications, putting your research at risk.

Ultrapure water (UPW), is water that has been purified to high standards. The water has a minimum H20 content and an equal amount of H+ and OOH-ions. It has a resistance of 18.2 MO.cm and TOC 10ppb. The bacterial count is 10 CFU/ml. Water must not contain any endotoxins to be considered ultrapure. This makes water a great reagent for laboratory research. Ultrapure water (also known as Type 1 water) reaches the theoretical optimal levels of purity. It has a resistivity value of 18.2 MO.cm and TOC values of 10 ppb. The bacterial count is 10 CFU/ml.

Endotoxins are removed and ultrapure water, also known as Type 1 Water, typically has a 0.03 EU/ml level with non-detectable nucleases or proteases.

Ultrapure water is essential for many sensitive scientific applications such as HPLC, LCMS, GCMS, GFAAS and mammalian cell cultivation, as well clinical analyzers.

Ultrapure Water and 2D Gel Electrophoresis:

Klose and O’Farrell developed two-dimensional gel electrophoresis (or 2D PAGE) independently in 1975. It is used to separate proteins from mixtures and has been a key method in protein analysis (proteomics). The 2D process consists of two separation principles. In the first dimension, ampholytes and immobilizing pH gradients are used to separate proteins according their isoelectric point. However, proteins in the second dimension are separated according their molecular weights using SDS PAGE. The separated proteins can then be visualized with different staining methods (Coomassie blue or silver staining). The separated proteins are then used to perform downstream analysis as required. There are many factors that can affect the quality of 2D gel electrophoresis results: The type and purity of the chemicals used, as well as the electrophoresis system. The advantages of horizontal high-performance electronphoresis (HPE), are well known. The advantages of using a horizontal high-performance electrophoresis (HPE) system over a traditional vertical system have been demonstrated. This study however examines the impact of water quality on silver staining. Two different grades of water were tested for their suitability.

Ultrapure water produced by the Sartorius Arium^® Pro VF water purification system

Water from the same system that had been stored for one week in a glass bottle (Erlenmeyer flask made of Duran glass and supplied by Schott; covered by perforated parafilm) at room temperature in a light place (referred to in this paper as “flask water”). The ultrapure water used was prepared as follows: The Arium^® Pro VF system was used to generate ultrapure water for 2D gel electrophoresis. It removes impurities still present in pretreated tap water.

Production of ultrapure water requires continuous recirculation and a constant water flow, which is achieved by a pump system with pressure control. The conductivity of the water is measured at the feed water inlet and in the product water (ultrapure water dispensed directly from the water outlet). The Arium^®, Pro VF, a predecessor model that was used in the above studies, has identical technical specifications to produce ultrapure water. It can be used with two different cartridges. They are filled with a mixed bed ion-exchange resins and an active carbon absorber to produce ultrapure water that has a low TOC (total organic carbon). A UV lamp, which can produce oxidizing or germicidal effects at wavelengths between 185 and 254 nanometers, is also integrated into the system.

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A profile of Sartorius

The Sartorius Group is a leading international partner of life science research and the biopharmaceutical industry. With innovative laboratory instruments and consumables, the Group’s Lab Products & Services Division concentrates on serving the needs of laboratories performing research and quality control at pharma and biopharma companies and those of academic research institutes. The Bioprocess Solutions Division with its broad product portfolio focusing on single-use solutions helps customers to manufacture biotech medications and vaccines safely and efficiently. The Group has been annually growing by double digits on average and has been regularly expanding its portfolio by acquisitions complementary technologies. In fiscal 2020, the company earned sales revenue of some 2.34 billion euros. At the end of 2020, nearly 11,000 people were employed at the Group’s approximately 60 manufacturing and sales sites, serving customers around the globe