An Easy-to-Construct Automated Winkler Titration System

The instrument described in this report is an updated version of the high precision, automated Winkler titration system described by Friederich et al.(1984). The original instrument was based on the work of Bryan et al. (1976) who developed a colorimetric endpoint detector and on the work of Williams and Jenkinson (1982) who produced an automated system that used this detector. The goals of our updated version of the device described by Friederich et al. (1984) were as follows: 1) Move control of the system to the MS-DOS environment because HP-85 computers are no longer in production and because more user-friendly programs could be written using the IBM XT or AT computers that control the new device. 2) Use more "off the shelf" components and reduce the parts count in the new system so that it could be easily constructed and maintained. This report describes how to construct and use the new automated Winkler titration device. It also includes information on the chemistry of the Winkler titration, and detailed instructions on how to prepare reagents, collect samples, standardize and perform the titrations (Appendix I: Codispoti, L.A. 1991 On the determination of dissolved oxygen in sea water, 15pp.). A disk containing the program needed to operate the new device is also included. (pdf contains 33 pages

Biopharmaceutical Process – Contract Development Organization: Startup

Due to their high specificity and the wide range of treatments they can provide, monoclonal antibodies (MAbs) from mammalian cell cultures have gained increasing popularity in therapeutics. As a result, treatments have become cheaper and easier to manufacture while maintaining their natural effectiveness, further increasing their appeal. Building MAb manufacturing facilities can be costly for biopharmaceutical companies, especially smaller biotech firms, and current production capacities are limited. As a result, there is an everincreasing demand for contract development organizations (CDOs). The CDO being proposed targets demand within this regime specific to MAbs entering clinical trials. It has the capability to screen clones, grow MAb-producing cells up to a 2500 L culture, and purify the MAb to clinical standards. By employing the newest technology available, the facilities will provide flexibility necessary for producing a myriad of different MAb therapeutics in Chinese Hamster Ovary (CHO) cells. Microbioreactors can screen dozens of clones at the millileter scale, saving time and money. Disposable bioreactors in the upstream process allow for variance in the production capacity due to the range of sizes they are available in. Finally, the purification process has been designed to allow for flexibility depending on the size and needs of every client’s product to maximize value to the costumer as well as the company. The current market for MAb production has an astounding worldwide value of approximately $27.5 billion and continues to expand as the number of MAbs entering clinical trials increases (Cowen 2006). It is estimated that within the next four years that the worldwide market value will reach$50 billion (“Preclinical Development”, 2010). The profitability of this proposal is based on running 39 batches a year at 4.326 kg MAb/batch or 168.71 kg MAb/year. By charging a reasonable average of $1,125,000/kg MAb, a profitability profile can be created. Assuming a 70$111,907,800 and 52.96% respectively. However, using a 70% production capacity also leaves room for even higher profit margins. The plant design also has space allotted for future expansion within the mammalian suite as well as room for a future microbial suite

Guidelines for the use of cell lines in biomedical research

Cell-line misidentification and contamination with microorganisms, such as mycoplasma, together with instability, both genetic and phenotypic, are among the problems that continue to affect cell culture. Many of these problems are avoidable with the necessary foresight, and these Guidelines have been prepared to provide those new to the field and others engaged in teaching and instruction with the information necessary to increase their awareness of the problems and to enable them to deal with them effectively. The Guidelines cover areas such as development, acquisition, authentication, cryopreservation, transfer of cell lines between laboratories, microbial contamination, characterisation, instability and misidentification. Advice is also given on complying with current legal and ethical requirements when deriving cell lines from human and animal tissues, the selection and maintenance of equipment and how to deal with problems that may arise

RRS Discovery Cruise 368, 15 Jul - 04 Aug 2011. Hydrographic measurements on WOCE line A16N

RRS Discovery Cruise 368 was a repeat occupation of part of the Atlantic hydrographic section designated by the World Ocean Circulation Experiment (WOCE) as A16N. A total of 29 CTDO (conductivity-temperature-depth-oxygen) stations were occupied. This included one test station, 27 stations between 49N and 23N on the WOCE A16N ‘20W’ line, and one final station near the ESTOC site close to Tenerife. Continuous profile measurements were CTDO and Lowered Acoustic Doppler Current Profiler (LADCP). Discrete bottle measurements from a 24-place rosette included salinity and dissolved oxygen analysed on board, and dissolved inorganic nutrients, Dissolved Inorganic Carbon and Total Alkalinity for analysis ashore. Underway measurements included Vessel-Mounted ADCP, surface ocean measurements and surface meteorology. The cruise was a UK contribution to the GO-SHIP sustained hydrography program. It was a partial repeat of the line designated in WOCE as A16N, which was previously occupied as a comprehensive cruise in 2003. In addition, a microbial program was carried out as an opportunistic activity by scientists who would remain on board for the following cruise.This report describes the methods used to acquire and process the data on board the ship during RRS Discovery Cruise 368.<br/

RRS James Cook Cruise 30, 26 Dec 2008-30 Jan 2009. Antarctic Deep Water Rates of Export (ANDREX)

This report describes scientific activities on RRS James Cook cruise 30, “ANDREX”, westwards from 30°E and in the vicinity of latitude 60°S, between late December 2008 and late January 2009. The cruise was terminated about halfway through by a medical emergency. Hydrographic work comprised 27 CTD/LADCP stations. Water samples were captured for measurement of salinity, dissolved oxygen, inorganic nutrients, oxygen isotope fraction, chlorofluorocarbons and sulphur hexafluoride, dissolved inorganic carbon and alkalinity, helium / tritium / noble gases and radiocarbon. Underway measurements comprised navigation, currents (ADCP), meteorology, and sea surface temperature and salinity. The remainder of the hydrographic section was executed a year later on RRS James Clark Ross, cruise JR239

Novel Low Shear 3D Bioreactor for the Scaled Production of High Purity Human Mesenchymal Stem Cells

Human mesenchymal stem cells are an ideal candidate for stem cell therapies. They have been researched since the 1960’s and can differentiate into many desired functional cell types without undergoing teratogenesis. However, higher yields are needed for a marketable, successful stem cell therapy. To accomplish this, cells will have to be cultured to expand them to therapeutically relevant dosages for multiple patients. Bioreactor production is an ideal method to solve this problem. The aim of this thesis is to test and validate a novel bioreactor for the cultivation of human mesenchymal stem cells. In this work, we investigate a novel suspended matrix for the culture on human mesenchymal stem cells (hMSCs). Initially we investigated various fiber meshes, both random and structured, for stem cell growth and morphology. We also investigated hMSC proliferation on rigid polymers commonly used in 3D printing. We then took the conditions that worked best in 2D culture and tested them in a small-scale model of the Express bioreactor from Sepragen. We have assessed cell growth on 3D printed Polylactic Acid (PLA) matrices and developed a scale down model bioreactor for development and characterization. Computational Fluid Dynamic (CFD) modeling was used in parallel with the described in-vitro experimentation to characterize shear profiles. From the CFD we were also able to predict a flow rate which resulted in almost zero shear. What we found was that hMSCs readily form confluent monolayers on the PLA lattice, and retain their surface marker expression and stemness. When combined with a short hypoxic treatment, the cells performed better than control flasks, resulting in a four-fold increase from seed with no impact on biomarker profile and differentiation ability