46
November/December 2009
Christopher Kirton (Huntington Life Sciences) followed with a talk entitled In vitro cytokine release’ posing the key question is this novel antibody therapeutic going to induce cytokine release syndrome (CRS)? This has been seen with antibodies such as Rituximab and more recently TGN1412, leading to release of TNF-, IFN- and IL-6, which can lead to serious adverse events. A bead- based muliplex immunoassay using flow cytometry detection has been set up and early results were presented at this meeting showing its potential to as a tool to predict CRS risk of a novel drug in vivo.
Small molecule re-analysis was picked up in the session too continuing the theme of the importance of this topic throughout the meeting with Jaap Wieling (Xendo) talk entitled ‘Accuracy of incurred samples’. To complete the session before lunch David Perrett (Queen Mary, University of London) gave a rather gruesome demonstration of the inadequacy of
sterilisation/decomtamination procedures in use in hospitals and dentists. Current procedures do not lead to the complete removal of Prion Proteins (PrP), the cause of new variant Creutzfeldt-Jakob disease (CJD). Even at very low levels PrP present a risk of cross-infection. Prion proteins are highly hydrophobic and have a high affinity for stainless steel, the material used for most surgical instruments. However new reagents are being developed and patented at Queen Marys to reveal contaminant PrPs in surgical instruments which will significantly reduce risk of cross-infection.
Session 8 – Regulatory approaches to evolving technologies (Chaired by Steve Westwood)
Brian Booth (FDA) joined the meeting by teleconference and covered (some of) what the FDA is thinking offering a personal view on current bioanalytical guidelines. Amongst the take home messages from Brian were that some issues of method validation are clear cut, while others depend on the situation. How much validation is required really does depend on the particular goal and everyone has a responsibility to ensure that the method is fit for its ultimate purpose. Brian concluded with the comment that the FDA can’t (won’t) figure everything out.
An impromptu discussion was held covering the 2008 EMEA Concept Paper on Bioanalytical method validation including Graeme Smith and Howard Hill (HLS) and Ray Briggs. Overall there were mixed views about the wisdom of ‘another’ set of guidelines which may lead to lack of clarity about validation requirements, however some delegates felt that having EMEA guidance would push the field forward, and challenge the FDA, which would be welcomed. The final session of the day chaired by John Lough (University of Sunderland) stayed on a large molecule theme with Jaap Weiling (Xendo) and Matt Ewles (Covance) presenting on immunoassays with elemental tagged antibodies and an introduction to the Quantification of proteins using proteolytic digestion by LC-MS/MS.
Session 9 – MS of Biological analytes (Chaired by John Smeraglia)
John Smeraglia (Pfizer) chaired this session with three talks given by Eric Ezan (CEA Institute of Biology and Technology), Chris Barton (Quotient Bioresearch) and
Figure 5. Slide from Brian Boothe’s presentation on ‘Bioanalytical Method Validation: (some of) What the FDA is thinking’.
Hendrick Neubert (Pfizer). The theme of this session was the application of LC- MS/MS to proteins, and its potential to supersede traditional immunoassays. Eric lead with a side-by side comparison of ELISA vs LC-MS. The initial challenge laid down by
Eric was that immunoassays are fundamentally flawed due to problems with lack of concordance of standards across platforms and questions the specificity of antibodies (what is measured? Free, bound or total analyte?). Therefore it is essential we look for alternatives. Could mass spectrometry be the answer?
Eric made a strong case for mass spectrometry as an alternative in the future particularly for biotherapeutics and in immunogenicity testing, linking in well with some of the challenges presented by Geoff Hale in earlier session. Further evidence was presented by Chris where the application of BioMS to biomarker testing has crossed species from equine testing looking at markers of drug abuse in horses, driven by the absence of suitable immunoassay methods) to the clinic where human samples are now being tested.
Hendrik brought the whole story together presenting his work on the development of an assay for low abundance targets of antibody therapeutics using immunoaffinity enrichment and LCMS-MS. This work demonstrates the power of combined approaches to answer complex sophisticated questions about percentages of free and bound drugs.
Session 10 – Concluding session (Chaired by Robin Whelpton)
In this final session John Stobaugh (University of Kansas) presented work on the evolution of chemical reactions for determination of protein 3-nitro-tyrosine, followed by Gavin O’ Connor (LGC) talking about protein quantification of mass spectrometry.
Conference closing comments John Lough (President of the Chromatographic Society) closed the conference commenting that it had been a good conference from a scientific standpoint with a wide variety of topics covered. Attendance was very good taking the present economic climate into consideration. (although down by around 20-30% on previous years). John thanked the organising committee and major sponsors of the event (Quotient Bioresearch Ltd, Waters and York Bioanalytical solutions).
Page 1 |
Page 2 |
Page 3 |
Page 4 |
Page 5 |
Page 6 |
Page 7 |
Page 8 |
Page 9 |
Page 10 |
Page 11 |
Page 12 |
Page 13 |
Page 14 |
Page 15 |
Page 16 |
Page 17 |
Page 18 |
Page 19 |
Page 20 |
Page 21 |
Page 22 |
Page 23 |
Page 24 |
Page 25 |
Page 26 |
Page 27 |
Page 28 |
Page 29 |
Page 30 |
Page 31 |
Page 32 |
Page 33 |
Page 34 |
Page 35 |
Page 36 |
Page 37 |
Page 38 |
Page 39 |
Page 40 |
Page 41 |
Page 42 |
Page 43 |
Page 44 |
Page 45 |
Page 46 |
Page 47 |
Page 48 |
Page 49 |
Page 50 |
Page 51 |
Page 52
