BIOSEPRA PROCESS PROTEOMICS SERVICES PDF

Broadly speaking, process omics refers to the application of molecular analytical tools to improve the manufacture of biological products. Suitable points at which this definition applies are also quite open-ended. While one would prefer analysis to occur during the process to provide feedback for optimizing process parameters on the fly, this will not become common practice among biotechnology companies for several more years, if ever. For now we must be satisfied with discovery-through-packaging as the relevant timeframe for applying the term process omics. Allison described methods that apply both genomics and proteomics to identify cellular pathways associated with high productivity. These pathways can then be manipulated to improve protein production, for example, by over-expression or knockdown experiments, or employed as biomarkers for higher productivity during process optimization.

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Broadly speaking, process omics refers to the application of molecular analytical tools to improve the manufacture of biological products.

Suitable points at which this definition applies are also quite open-ended. While one would prefer analysis to occur during the process to provide feedback for optimizing process parameters on the fly, this will not become common practice among biotechnology companies for several more years, if ever. For now we must be satisfied with discovery-through-packaging as the relevant timeframe for applying the term process omics.

Allison described methods that apply both genomics and proteomics to identify cellular pathways associated with high productivity. These pathways can then be manipulated to improve protein production, for example, by over-expression or knockdown experiments, or employed as biomarkers for higher productivity during process optimization. Similarly, proteomics may be employed to characterize protein expression productivity as a result of genomic manipulations or in response to changes in culture medium.

Among proteomic approaches, Dr. Allison described traditional and emerging gel electrophoretic methods as well as more advanced mass spectrometric measurements. One emerging omics technique, metabolomics, measures panels of small molecule biomarkers for almost any cellular process, either native or in response to process changes. Metabolomics is not as simple as measuring levels of a protein product.

Metabolomic information must be correlated to the primary measure gene, protein and often extrapolated to be useful. Since protein production is the objective, proteomics are the most useful.

SELDI mass spectrometry acts as a rapid, straightforward mass detector for proteins ideal for optimizing separation systems, says Pall. At the time of the Biosepra acquisition, Pall announced plans to establish process proteomics service centers to help customers select and optimize sorbents and membranes for protein purification. Pall claims that microscale methods and processes are scalable to pilot and manufacturing scale. Invitrogen www. PD-Direct operates from cell line selection through media development to scale-up for manufacturing.

Carrier says. Programs like PD-Direct are causing Invitrogen to rethink how genomics and proteomics fit into the overall development scheme, particularly in cell-line development. Most process omics efforts to date, notes Dr. Carrier, have been based on cellular mechanisms. Invitrogen is working on a number of contracts using PD-Direct Revolution to improve existing production cell lines for products entering early clinical studies.

Revolution is for proteins that are difficult to express, are not secreted, or are inhibitory to cells. Incorporating omics technologies during development and manufacturing is an ongoing, evolving effort at Invitrogen, Dr. Carrier admits. As process omics evolves, vendors will increasingly seek to leverage their expertise in laboratory and discovery-stage instrumentation to process settings.

Beckman Coulter www. Since then, the company expanded ProteomeLab further by drawing on its expertise in chromatography and capillary electrophoresis to provide proteomic analytical systems suitable for discovery through production. Among these systems is the PA for automated, high-resolution protein characterization and the PF 2D automated 2-D protein fractionation system.

Primarily a discovery tool, PF 2D employs chromatofocusing in the first separation dimension and reverse phase in the second. According to Mark Lies, Ph. Protein identities are then confirmed through mass spectrometry. The system, which has been widely adopted by the biotech industry as a QC tool, is packaged with off-the-shelf assays, chemistries, and protocols. Included chemistries include CE-SDS a replacement for SDS-polyacrylamide gel electrophoresis for molecular weight separation, capillary isoelectric focusing separation based on isoelectric point and charge heterogeneity , and carbohydrate profiling, essentially a microheterogeneity assay.

The system looks at protein purity, heterogeneity, and ultimately, identity. Process omics has its most profound impact not on processing or in-process analytics, but during cell-line selection, clonal expansion, and the selection and development of cell culture media.

Many cell culture experts, most notably Florian Wurm, Ph. Wurm has developed high-throughput technology to optimize and compress development times for media through a more-or-less traditional approach: altering one or more variable s , measuring protein output, and analyzing results statistically. CellSpot provides direct, simultaneous readouts on individual cells for multiple characteristics, for example protein secretion, surface proteins, and intracellular signal transduction pathways.

Traditionally, cell-line development occurs through screening hundreds to thousands of individual cell culture supernatants. By taking the pooled average for all the cells in the culture, this method misses the individuality of cells within heterogeneous populations. CellSpot assays the productivity of up to millions of cells simultaneously, quantitatively, and individually after minimal cell growth periods.

Supernatant monitoring requires much longer growth. These advantages lead to significant reduction in cell-line development times, by up to half, for both library screening and sub-cloning. For the latter, CellSpot sensitivity is adjusted to account for smaller productivity differences among cells.

CellSpot combines nanoparticle-based detection, automated digital microscopy, and software to make sense out of protein-secretion data. The polystyrene nanoparticles, measuring — nm in diameter, are coated with an antibody to the target protein and a dye to trigger the light emission used for detection, similar to a sandwich ELISA assay.

CellSpot works for all mammalian cells, adherent or nonadherent, and expression systems, according to Dr. Cell libraries are plated on a surface modified to capture the antibody or recombinant protein secreted by individual cells located within virtual wells approximately microns across.

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Omics Prove Handy in Production Processes

The BioSepra Process Division develops, manufactures and markets chromatography sorbents that greatly simplify protein purification for drug development and production. Eric Krasnoff, Chairman and CEO of Pall Corporation, said, "The addition of this innovative, broad range of chromatography chemistries strengthens our ability to serve the needs of biotechnology customers. The marriage of BioSepra's capabilities and Pall's extensive existing product portfolio allows customers to benefit from a highly integrated 'Total Fluid Management' approach. Danaher To Acquire Pall Corporation. These chromatographic systems enable single-use, multicolumn chromatography for process development and are well placed for multiproduct and single-use facilities. Pall Corporation launches a comprehensive line of chromatography sorbents. These advanced industry-proven sorbents from Pall greatly simplify protein separation and purification for biotechnology drug discovery, development and production.

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Pall Acquires Chromatography Business from Ciphergen

Exhibit For Immediate Release:. Ciphergen Biosystems, Inc. Sue Carruthers. Investor Relations The transaction is expected to close within 45 days, subject to customary conditions of closing.

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Pall completes protein purification acquisition

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