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RAPID Bioanalytical METHODS

Research Goals Current Projects Results Publications Collaborations & Funding

Research Goals

The primary goal of the research is concerned with developing rapid, quantitative spectroscopic based techniques for the non-contact, non-destructive analysis of complex materials used in biological based pharmaceutical manufacturing.

The value of the methods that we are developing lies in the fact that they are inexpensive and suitable for screening all the materials used in a BioPharmaceutical manufacturing. These methods are designed to be very rapid, require no expensive training or consumables and are also perfectly suited to use in aseptic methods of production and harsh environments.

To date we have focused on the quantitative analysis of cell culture media, the raw materials used for medias, and bioprocess broths. We have been provided with unique access by our industrial partner to a complete production cycle of samples, 30 times over.  This has enabled us to build very robust and rapid analytical tools suitable for all stages of production. 

Jan. 11: Our work on rapid cell culture media analysis has been featured in the January issue of Genetic Engineering & Biotechnology News.  The article can be accessed here.

CBAS (Phase 1, BMS), 2005-2009:

A team of ~10 researchers was based in the Nanoscale Biophotonics Laboratory on this project. The CBAS project was a collaboration with Bristol-Myers Squibb and Dublin City University and is funded by the IDA and BMS.
Press releases on the project can be found on the Galway Advertiser, and IDA website.
A discussion piece is also available on: PharmaManufacturing.Com. 

This project is now complete, although we are still publishing the outcomes of this research, and we hope to have several new papers in 2012-13.

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Current Projects

We are currently undertaking a wide variety of individual research projects.  Details will be posted in the near future, however much of what we do is covered by confidentiality agreements.  In general, the work involves the use of Raman, FT-IR, FT-NIR, and Fluorescence spectroscopies for the quantitative analysis of complex mixtures. The research involves the development of novel chemometric approaches for data analysis, coupled with spectroscopic methods.

Phase II, 2010+

A new industrial collaboration with the Janssen Pharmaceutical Companies of Johnson & Johnson began on the 01/09/10.  The project, funded by J&J  and NIBRT developed new methods for characterizing cell culture media.

Phase III, 2011-12.

We have just begun a collaboration with Merck on the analysis of cell culture media and the development of new analytical methods.This research involves the use of Raman and Fluorescence spectroscopies.

 

Cell culture media: 

Mammalian cell culture is increasingly being used for the manufacture of recombinant protein based therapeutics. One of the key components of the cell culture manufacturing process is the area of cell culture media (CCM).  The fields of CCM formulation, storage, feeding, and analysis is a critical area of bio-analytical science which is complicated by the fact the CCMs are very complex and sensitive materials, particularly when in liquid form.   CCMs can consist of a wide variety of diverse components: sugars, amino acids, proteins, vitamins, inorganic salts and other nutrients. However the rapid, online, non-destructive, and quantitative analysis of CCMs is currently and under-developed area, but there is an urgent need for these types of methods. Rapid and/or real-time monitoring of CCM composition would provide a tool to improve process control and product yield. To optimize the product yield, the CCM composition and feeding rates must be tightly controlled.

Motivated by the need for rapid, non-contact, non-destructive and inexpensive methods of analysis for these complex CCM mixtures, we are exploring the application of Raman and Fluorescence spectroscopy coupled with mathematical analysis for robust, reliable, and rapid CCM characterization.

Bioprocess broths:

The analysis of complex cell culture process broths is challenging, time-consuming, and expensive due to the fact that they are complex mixtures  and consist of a vast variety of components, e.g., inorganic salts, sugars, amino acids, proteins, vitamins, and other nutrients. This thus requires the development of holistic and inexpensive methods that can elucidate compositional information from these complex mixtures for diagnostic and process control purposes. Here we have used a range of spectroscopic methods coupled with advanced Chemometrics to develop a suite of characterization tools which are all suited to online process control. 

BioPharma raw materials:

A lot of the raw materials or third party supplied media components are very complex mixtures (solid or liquid) and they are challenging from an ID and quality assessment point of view. We have developed a range of very inexpensive ID and quality tools for use in both the QC lab and in the storeroom. 

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Selected Results

We will upload data when appropriate. We expect to be able to publish a range of manuscripts detailing our work over the past 5 years, once reviewed by our industrial partners.  

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Publications and Conference Presentations

More details, posters, and manuscripts will be uploaded as they become available and as they are passed by the BMS IP review committee.

Publications (updated 26-09-2012): 

1. Rapid Quantification of Tryptophan and Tyrosine in Chemically Defined Cell Culture Media using Fluorescence Spectroscopy. A. Calvet, B. Li, and A. G. Ryder.  Journal of Pharmaceutical and Biomedical Analysis, 71, 89-98, ( 2012). DOI: 10.1016/j.jpba.2012.08.002
2. Using Surface-Enhanced Raman Scattering (SERS) and Fluorescence Spectroscopy for Screening Yeast Extracts, A Complex Component of Cell Culture Media. B. Li, N.M.S. Sirimuthu, B. Ray, and A.G. Ryder,   Journal of Raman Spectroscopy, 43(8), 1074-1082, ( 2012).
   DOI:  10.1002/jrs.3141
   
3. Fluorescence EEM Spectroscopy for Rapid Identification and Quality Evaluation of Cell Culture Media Components.  B. Li, P.W. Ryan, M. Shanahan, K.J. Leister, and A.G. Ryder. Applied Spectroscopy, 65(11), 1240-1249, ( 2011). 
   DOI:  http://dx.doi.org/10.1366/11-06383.
4. Rapid, low-cost, quantitative characterization of cell culture media using a combination of spectroscopic and chemometric methods.  A.G. Ryder, B. Li, and K.J. Leister.  International Drug Discovery, Oct./Nov. issue, 18-21, ( 2010).
   
5. Rapid Characterization and Quality Control of Complex Cell Culture Media Solutions using Raman Spectroscopy and Chemometrics. B. Li, P.W. Ryan, B.H. Ray, K.J. Leister, N.M.S. Sirimuthu, and A.G. Ryder.  Biotechnology and Bioengineering, 107(2), 290-301, ( 2010).
   DOI: 10.1002/bit.22813 .
   
6. Prediction of Cell Culture Media performance using Fluorescence Spectroscopy.  P.W. Ryan, B. Li, M. Shanahan, K.J. Leister, and A.G. Ryder, Analytical Chemistry, 82(4), 1311-1317, ( 2010).
   DOI: 10.1021/ac902337c  Available online now at: http://pubs.acs.org/doi/abs/10.1021/ac902337c
   
7. A Stainless Steel Multi-Well Plate (SS-MWP) for High Throughput Raman Analysis of Dilute Solutions. A.G. Ryder, J. de Vincentis, B. Li, P.W. Ryan, N.M.S. Sirimuthu, and K.J. Leister.   Journal of Raman Spectroscopy, 41(10), 1266-1275, ( 2010).  DOI: 10.1002/jrs.2586.

Another tranche of manuscripts are under review and several more are in preparation for submission to the CBAS IP committee for review by the middle of 2013. 

Relevant Presentations (as of 24-07-2012): 

1. Quantitative analysis of cell culture media using multi-dimensional fluorescence spectroscopy, A.G. Ryder,  Biopharmaceutical Raw Materials: Qualification, Analysis, & supply chain management, Cologne, Germany, 27-28 June, 2012 (invited).
2. Quantitative Analysis of Cell Culture Media using Raman Spectroscopy. A. Ryder,  26th  International Forum and Exhibition Process Analytical Technology,  Baltimore, MD, USA, 22-25 Jan. 2012 (invited).
3. Rapid, Quantitative analytical methods for hydrosylates using in cell culture media:  a spectroscopic approach.  Alan Ryder,  Process2Product, Translating Process Knowledge into Quality Biological Products, Washington D.C., USA, 4-6 October 2011 (invited).
4. Quantification of amino acids and vitamins in cell culture media using Excitation-Emission Matrix fluorescence spectroscopy and chemometrics.  A. Calvet and A.G. Ryder, 12th Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging and Probes, Strasbourg, France, 11-14 Sept., 2011.
   
5. Prediction of bioprocess performance by multidimensional fluorescence spectroscopy analysis of bioreactor broths.  B. Li and A.G. Ryder, 12th Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging and Probes, Strasbourg, France, 11-14 Sept., 2011.
   
6. Quantitative analysis of yeastolate and eRDF in model cell culture media using fluorescence spectroscopy.  B. Kissane, B. Li, and A.G. Ryder, 12th Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging and Probes, Strasbourg, France, 11-14 Sept., 2011. 
   
7. Quantitative Analysis of Yeastolate and eRDF in aqueous mixtures by Surface Enhanced Raman Scattering (SERS).  B. Kissane, B. Li, N.M.S. Sirimuthu, and A.G. Ryder. RSC Analytical Research Forum 2011, University of Manchester, UK, 25 - 27 July 2011.
   
8. 2D fluorometry to monitor CHO cell based bioprocesses used in biopharmaceutical manufacturing.  B. Li and A. G. Ryder. RSC Analytical Research Forum 2011, University of Manchester, UK, 25 - 27 July 2011.
   
9. Rapid, Low-Cost, Quantitative Characterization of Cell Culture Media Using a Combination of Spectroscopic and Chemometric Methods.  Alan Ryder,  Process2Product, Translating Process Knowledge into Quality Biological Products, Bethesda, Maryland, USA, 21-22 Oct., 2010 (invited).
   
10. Component Tracking of Complex Aqueous Cell Culture Media Using Multidimensional Fluorescence and Chemometric Methods.  B. Li, A.G. Ryder, P.W. Ryan, and K.J. Leister.  11th Conference on Methods and Applications of Fluorescence: Spectroscopy, Imaging and Probes, Budapest, Hungary, 6-9 Sept., 2009.
    
11. Determination of glucose concentration in cell culture media by Raman spectroscopy and multivariate calibration. B. Kissane, A.G. Ryder, & B. Li, 42nd IUPAC Congress, Glasgow, 2-7 Aug. 2009.
    
12. Characterisation of hydroxyapatite and dicalcium phosphate using X-Ray Fluorescence Spectrometry, N. Walshe, A.G. Ryder, C. O'Kane, & A.R. Boyd, 42nd IUPAC Congress, Glasgow, 2-7 Aug. 2009.
13. Process monitoring of a complex aqueous cell culture broth by multidimensional fluorescence and chemometric methods. B. Li, A. G. Ryder, P.W. Ryan, & K.J. Leister, 42nd IUPAC Congress, Glasgow, 2-7 Aug. 2009. 
14. A strategy for rapid Raman characterisation and quality control of cell culture media components. P.W. Ryan, B. Li, A.G. Ryder, K.J. Leister, B.H. Ray, & N.M.S. Sirimuthu, 42nd IUPAC Congress, Glasgow, 2-7 Aug. 2009.
    
15. Surface enhanced Raman spectroscopy (SERS): a powerful tool to monitor the degradation of complex raw materials used for Biopharmaceutical Manufacturing. A.G. Ryder, N.M.S. Sirimuthu, and B. Li. International Conference on Trends in Bioanalytical Sciences and Biosensors ( ICTBSB-2009), Dublin, 26-27 Jan., 2009.
16. Methods for the Rapid Identification of Cell Culture Media Components. P.W. Ryan, B. Li, A.G. Ryder, K.J. Leister, and N.M.S. Sirimuthu. International Conference on Trends in Bioanalytical Sciences and Biosensors ( ICTBSB-2009), Dublin, 26-27 Jan., 2009.
17. Multidimensional fluorescence analysis of complex aqueous cell culture media by chemometric methods.  B. Li, A.G. Ryder, P.W. Ryan, and K.J. Leister. International Conference on Trends in Bioanalytical Sciences and Biosensors ( ICTBSB-2009), Dublin, 26-27 Jan., 2009.
18. Quantitative Analysis of Yeastolate and eRDF in aqueous mixtures by Surface Enhanced Raman spectroscopy (SERS).  B. Kissane, A.G. Ryder, B. Li, and N.M.S. Sirimuthu. International Conference on Trends in Bioanalytical Sciences and Biosensors ( ICTBSB-2009), Dublin, 26-27 Jan., 2009.
19. Eliminating the water signal from Raman spectra of aqueous samples. R. Rajko, A.G. Ryder, B. Kissane, and B. Li.   Metabolomics Society 3 ^rd Annual Conference, Manchester, UK, 11-14 June 2007.
20. A novel similarity index for quality assessment of complex aqueous media by multidimensional fluorescence spectroscopy.  B. Li,  R. Rajko, B. Kissane, and A.G. Ryder.  Metabolomics Society 3 ^rd Annual Conference, Manchester, UK, 11-14 June, 2007.

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Collaborations & funding

We are happy to work with industrial partners in Europe and US who are interested in this type of spectroscopic approach to complex biotechnology processes.  For Bio-pharmaceutical companies based in Ireland:

• Enterprise Ireland offer a range of supports to leverage additional support for Industry-Academia partnerships.
  
• IRCSET also offer the possibility of jointly funded PhD scholarships under the Enterprise Partnership Scheme.
• SFI have a Technology and Innovation Development Award (TIDA) which can be used to support short 1 year, research projects. 

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