Throughput Optimization of Continuous Biopharmaceutical Manufacturing Facilities.

In order to operate profitably under different product demand scenarios, biopharmaceutical companies must design their facilities with mass output flexibility in mind. Traditional biologics manufacturing technologies pose operational challenges in this regard due to their high costs and slow equipment turnaround times, restricting the types of products and mass quantities that can be processed. Modern plant design, however, has facilitated the development of lean and efficient bioprocessing facilities through footprint reduction and adoption of disposable and continuous manufacturing technologies. These development efforts have proven to be crucial in seeking to drastically reduce the high costs typically associated with the manufacturing of recombinant proteins. In this work, mathematical modeling is used to optimize annual production schedules for a single-product commercial facility operating with a continuous upstream and discrete batch downstream platform. Utilizing cell culture duration and volumetric productivity as process variables in the model, and annual plant throughput as the optimization objective, 3-D surface plots are created to understand the effect of process and facility design on expected mass output. The model shows that once a plant has been fully debottlenecked it is capable of processing well over a metric ton of product per year. Moreover, the analysis helped to uncover a major limiting constraint on plant performance, the stability of the neutralized viral inactivated pool, which may indicate that this should be a focus of attention during future process development efforts.LAY ABSTRACT: Biopharmaceutical process modeling can be used to design and optimize manufacturing facilities and help companies achieve a predetermined set of goals. One way to perform optimization is by making the most efficient use of process equipment in order to minimize the expenditure of capital, labor and plant resources. To that end, this paper introduces a novel mathematical algorithm used to determine the most optimal equipment scheduling configuration that maximizes the mass output for a facility producing a single product. The paper also illustrates how different scheduling arrangements can have a profound impact on the availability of plant resources, and identifies limiting constraints on the plant design. In addition, simulation data is presented using visualization techniques that aid in the interpretation of the scientific concepts discussed.

External quality assessment of HbA1c and its effect on comparison between Swedish pediatric diabetes clinics. Experiences from the Swedish pediatric diabetes quality register (Swediabkids) and Equalis.

BACKGROUND: To explore to what extent measurement error can explain the variation of mean patient HbA(1c) between clinics. METHODS: For each year 2005-2010 data from 5380-6985 children, age <18 years, in 35-43 Swedish pediatric clinics was analyzed. Each year 13,000-19,000 HbA(1c) analyses were evaluated. Year mean HbA(1c) for each patient was calculated for HbA(1c) values when insulin dose was ≥0.5 U/kg. In Sweden HbA(1c) values were during the study period standardized to the Mono S level, HbA(1c)(Mono S)%, but are given also in the international unit HbA(1c)(IFCC), mmol/mol. Performance of locally measured HbA(1c) is monitored by Equalis through monthly external quality assessment (EQA) schemes. RESULTS: The yearly mean bias term for each clinic varied from -0.54 to 0.41 HbA(1c)(Mono S)%. The bias between clinic HbA(1c) and target value improved during the 6 years and the mean bias was for 79%-88% of clinics within the recommended level ±0.14 HbA(1c)% the last 2 years. Inter-clinic mean HbA(1c) had a wide interquartile range, 0.30-0.43 HbA(1c)(Mono S)% [3.2-4.5 HbA(1c)(IFCC)mmol/mol]. CONCLUSIONS: Regular participation in EQA schemes is necessary when comparing HbA(1c) values. The measurement error decreased during the 6-year period and explained from 28% to <10% of the inter-clinic variation in year mean clinic HbA(1c).

A fast and inexpensive procedure for the isolation of synthetic cannabinoids from 'Spice' products using a flash chromatography system.

In the age of the Internet, the variety of drugs offered online is constantly increasing, and new drugs emerge every month. One group of drugs showing such an enormous increase is that of synthetic cannabinoids. Since their first identification in 'herbal mixtures', new structural modifications continue to appear on the market. In order to keep up with this process, toxicological screening methods need to be up to date. This can become extremely difficult if no reference material is available. In this article, a fast and effective way to extract and purify synthetic cannabinoids from 'herbal mixtures' is presented. This method opens a new opportunity for a timely reaction by obtaining reference material straight out of the 'herbal mixtures' ordered via the Internet. Isolation was carried out on a flash chromatography system with gradient elution on a C18 column using methanol and 0.55 % formic acid as mobile phases. The obtained purity of all compounds exceeded 99 %. In addition to the isolation of single compounds, the method proved to be suitable for the separation of various synthetic cannabinoids in one mixture, including the diastereomers cis- and trans-CP-47,497-C8. This approach for obtaining pure standards of new drugs proved to be effective, inexpensive and much quicker than waiting for the substances to be commercially available as reference material.

New theory for distribution of minimum resolution in multi-component separations with noise/detection limits.

Equations were proposed recently for computing the distribution of minimum resolution (resolution distribution) of two Gaussian peaks with equal standard deviations, when peak heights in a multi-component separation follow a statistical distribution. The computation depended on the survival function of the peak-height ratio. Previously, an equation was derived for a first-order survival function that excluded peaks with heights less than a noise/detection limit. Here, an equation is derived for a corrected survival function, under the more realistic assumption that two minimally resolved peaks are lost if the height of their shoulder is less than the noise/detection limit. First-order and corrected survival functions and resolution distributions are derived for the exponential and uniform distributions of peak heights, and a corrected survival function and resolution distribution are derived for the log-normal distribution (LND) to complement a previous first-order derivation. Large peak losses (up to 99.3% of the noise/detection limit) are considered to find significant differences between the first-order and corrected resolution distributions. For the LND and exponential distribution, the corrected resolution distribution has slightly greater density in the low-resolution region but otherwise differs little from its first-order counterpart, unless the scale parameter of the LND is small (e.g. 0.75). For the uniform peak-height distribution, the corrected resolution distribution has higher density in the high-resolution region. The first-order and corrected resolution distributions are almost the same as long as the first moment of the first-order resolution distribution is greater than 0.6. The predictions are confirmed by Monte-Carlo simulation.

External quality assessment of multi-analyte chromatographic methods in oral fluid.

BACKGROUND: A proficiency testing scheme was set up for the DRUID (Driving under the influence of Drugs, Alcohol and Medicines) research project, funded by the European Commission, in which oral fluid is analysed by eleven laboratories. A common collection and analysis methodology is used: Statsure Saliva Sampler is used for collection and LC-MS/MS or GC-MS confirmation analysis of 22 substances is performed on all samples. Despite internal validation and quality control samples, external quality assessment is still necessary to further increase comparability of results. Four rounds of proficiency testing (PT) were organized between March 2008 and September 2009. METHODS: Qualitative results were evaluated using sensitivity and specificity. Quantitative results were evaluated using z-scores and the standard deviation of Horwitz. RESULTS: Specificity was above 99% in each round, sensitivity per analyte varied between 81.7 and 100%, and 20 out of 22 analytes had a sensitivity above 90%. The percentage of satisfactory z-scores increased from 79.4% to 89.2%. This trend was seen for all drug classes, except zopiclone. Results were discussed with participating laboratories and problems were addressed. CONCLUSIONS: Because of these corrective actions, DRUID laboratories have a lower variation in results than previously published PT schemes in oral fluid.

An effective method for hemoglobin E detection: DEAE sepharose microcolumn.

Hemoglobin E (Hb E) is a variant hemoglobin that can lead to considerable morbidity when it occurs in a compound heterozygous state with beta-thalassemia. Therefore, its detection is important because it permits antenatal counseling. Hb E is prevalent in economically weaker regions of the world. Thus, its recognition is often hindered by the high costs of sophisticated Hb E-detection assays. We developed a simple visual test based on identifying Hb E in blood samples with DEAE Sepharose microcolumn chromatography. The diagnostic accuracy of our new Hb E microcolumn assay was 100% for sensitivity, specificity, and positive and negative predictive values. Furthermore, the Hb E microcolumn assay is rapid, reproducible, and economical; hence, it offers affordable screening for Hb E in less well-equipped laboratories.

Disposables in downstream processing.

Disposable equipment has been used for many years in the downstream processing industry, but mainly for filtration and buffer/media storage. Over the last decade, there has been increasing interest in the use of disposable concepts for chromatography, replacing steel and glass fixed systems with disposable plastic modules that can be discarded once exhausted, fouled or contaminated. These modules save on cleaning and validation costs, and their reduce footprints reduce buffer consumption, water for injection, labor and facility space, contributing to an overall reduction in expenditure that lowers the cost of goods. This chapter examines the practical and economic benefits of disposable modules in downstream processing.