Author Archives: BPI Contributor

Bioreactor Monitoring – Best Practices for Cell Counting, Sizing, and Viability

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Manufacturers utilizing bioprocesses for production need accurate and reliable tools for monitoring bioreactors. The key parameter that many bioprocess engineers rely on is viable cell density. This is a compound parameter arising from knowledge of the cellular concentration and the total cellular viability. Viable cell density is the mathematical product of these two primary pieces of information. In order to minimize variability, the detection technique of each of the primary measurements (concentration and viability) must be understood separately.

In this educational webcast, Matthew N Rhyner, PhD., Global Analytical Marketing Manager at Beckman Coulter Inc, discusses two of the most powerful techniques for assessing concentration and viability: the Coulter Principle and flow-based imaging. Join Dr. Rhyner as he compares the strengths and weaknesses of these techniques and discusses competing technologies. The webcast concludes with some recommendations for best practices in monitoring cell count, size, and viability.

October 1, 2013

Viral Vector Production in the Integrity® iCELLis® Disposable Fixed-bed Bioreactor from Bench-scale to Industrial Scale

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Recombinant viruses (e.g. lentivirus and adeno-associated-virus) can be used as human gene therapy vectors. They are mainly produced in adherent cell cultures (e.g. HEK293T, A549, VERO) in Roller Bottles or multiple-tray-stacks using either transient transfection or infection strategies. Therefore, the Integrity® iCELLis® (ATMI LifeSciences) line of bioreactors offers a new production alternative with stronger process controls and ease of scale-up.   The iCELLis bioreactor is designed for adherent cell culture applications. Cells grow on microfibers carriers packed in a fixed-bed…

September 26, 2013

Is the Gold Standard Tarnished? Weaknesses in the Standard Method Compromise Your RMM Validation

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Introduction of a new Rapid Microbial Method (RMM) requires a full validation. The purpose of the validation is to prove that equivalent or better results are obtained using the new method when compared to the compendia. One assumes that the result obtained with the compendial method is absolute and is the “Gold Standard†by which to compare the new test. In reality, the compendial method exhibits a number of weaknesses that compromise the integrity of the validation. The standard requires…

September 13, 2013

Culture Media: Sourcing Animal-Free Raw Materials

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For September’s Ask the Expert, we talked to Bill Whitford (senior manager of the bioprocess market for Thermo Fisher Scientific) about sourcing animal-free raw materials for cell culture media. This has become a requirement for many biopharmaceutical sponsors, but it is not without its issues. But what precisely is meant by animal is not always clear. Regulatory guidance is sparse on this issue. And a designation of animal-derived–component free (ADCF) often involves additional certification, testing, or auditing. Sponsors and material…

September 1, 2013

Reducing Tech-Transfer Risk

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Process understanding is the key to mitigating risks associated with technology transfer. In 2007, Justin Neway provided strategies for combining on-demand data access, trending, reporting, and analytics to achieve that understanding throughout process development (pdf). With increasing adaptation of process analytical technology (PAT) and quality-by-design (QbD), process understanding strategies have shifted.

September 1, 2013

A Platform Approach for the Purification of Domain Antibodies (Dabs)

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A three-step purification process for a Dab successfully developed and verified. The step yield ranged between 86% and 99%, giving a total process yield of approximately 81%. The ECP in the start sample was more than 200 000 ppm whereas the final sample contained only 5.5 ppm. The endotoxin content in the feed was approximately 2 million endotoxin units/mg of protein and the final sample was below the limit of quantification. Protein L leakage was undetectable in all samples. This…

September 1, 2013

MFI for Particle Characterization of Biopharmaceuticals Today

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Micro-flow imaging (MFI) is a sensitive, simple and automated method for the analysis of sub-visible particles and translucent protein aggregates that provides particle size, count and morphology. Because it quantifies particle shape, this technology can discriminate groups of particles from each other, and evaluate how these groups change over time. In this article, we provide an overview of how biopharma is using MFI to characterize and monitor their protein formulations in new and emerging applications.  

September 1, 2013

POROS® CaptureSelect™ Affinity Columns for Rapid, Small-Scale Purification and Sample Preparation of Recombinant Proteins

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This application note describes a model system in which Enbrel® protein, a fusion protein consisting of TNF receptor and IgG1 Fc, is purified from a dilute sample matrix for further analytical characterization. The resulting purified protein was subjected to N-linked glycan structural analysis by mass spectrometry (MS) and its aggregation state was assessed by analytical ultracentrifugation (AUC). This data set serves as a model to demonstrate the capability of the POROS® CaptureSelectâ„¢ product line for the affinity purification of biomolecules…

September 1, 2013

Upstream Processing

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The “Cell Culture and Upstream Processing” track began many years ago as a conference of its own. In October 2004, IBC Life Sciences brought it together in Boston, MA, with three other events (“Recovery and Purification,” “Production and Economics,” and “Scaling Up from Bench to Clinic”) to create the first BPI Conference. Just like BPI magazine’s editors, the event producers have depended on industry advisors from the beginning. Our editorial advisory board (EAB) members give generously of their time and…

September 1, 2013

Downstream Processing

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The “Recovery and Purification” track began many years ago as a conference of its own. In October 2004, IBC Life Sciences brought it together in Boston, MA, with three other events (“Cell Culture and Upstream Processing,” “Production and Economics,” and “Scaling Up from Bench to Clinic”) to create the program of the first BPI Conference and Exhibition. Just like BPI magazine’s editors, the event producers have depended on industry advisors since the beginning. Our editorial advisory board (EAB) members give…

September 1, 2013