Novel use of accelerator mass spectrometry for the quantification of low levels of systemic therapeutic recombinant protein

Lappin, Graham; Garner, R. Colin; Meyers, Tunde; Powell, John; Varley, Paul

doi:10.1016/j.jpba.2006.02.019

Cited by 19 publications

(41 citation statements)

References 9 publications

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“…However, with the exception of two articles [10,19] all data relate to conventional drugs with molecular weights in the 100-500 Da range. Larger molecules including proteins and polysaccharide back-bone drugs are of great interest within the pharmaceutical industry.…”

Section: Macromolecular Pharmacokinetic Studiesmentioning

confidence: 99%

Accelerator mass spectrometry of ultra-small samples with applications in the biosciences

Salehpour¹,

Håkansson²,

Possnert³

2013

Nuclear Instruments and Methods in Physics Research Section B:

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Section: Macromolecular Pharmacokinetic Studiesmentioning

confidence: 99%

Accelerator mass spectrometry of ultra-small samples with applications in the biosciences

Salehpour¹,

Håkansson²,

Possnert³

2013

Nuclear Instruments and Methods in Physics Research Section B:

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“…Similar differences have been seen by the author in other studies where AMS and LC/MS have been compared, as well as in studies where AMS has been compared with ELISA. 55 The studies described in this section all support the concept that AMS has utility in measuring intracellular drug concentrations. It is interesting to speculate what other areas of drug development might lend themselves to intracellular drug concentration measurement.…”

Section: Microdosing and Drug Targetingmentioning

confidence: 52%

Human Microdosing/Phase 0 Studies to Accelerate Drug Development

Garner¹

2014

Human-Based Systems for Translational Research

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There is an urgent need to accelerate the development of new drugs to treat unmet medical needs both in the developed and underdeveloped parts of the world. There is also an urgent need to improve the efficiency of drug development through time and cost reduction because development costs hinder the introduction of new therapies. There are many life-threatening human diseases that still require better therapies including cancer, bacterial and viral infection and lifestyle-associated diseases. The costs of bringing a drug to market are so high that there are very few organisations able to afford to discover, develop and introduce drugs on their own. Estimates of drug development costs vary between US$1.2 and 12.0 billion dollars, making the costs of newly approved drugs very high in order that companies can recover the costs of all the failed drugs in their pipelines. 1,2 It has been estimated that only three out of 10 drugs make a return on their investment and that over 99% of molecules researched by a pharmaceutical company are likely to fail at some point during the development process. 3 Even when a drug enters into RSC Drug Discovery Series No. 41 Human-based Systems for Translational Research Edited by Robert Coleman

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“…31 Measurement of protein therapeutics in animals and humans is difficult, as analytical methods such as immunoassays frequently lack sensitivity and are subject to interferences. 12,30 Analysis is further complicated by the fact that many protein drugs are nearly indistinguishable from proteins that occur naturally in the body. 12,31 …”

Section: Discussionmentioning

confidence: 99%

“…9–11 In addition, measurement of protein therapeutics in experimental animals using 14 C-AMS has recently been reported. 12,13 , 14 C-AMS is an attractive approach for quantitation of therapeutic proteins or low-abundance post-translational modifications displayed on proteins because its sensitivity allows the use of doses that impose negligible chemical and radioactive risk to humans, while providing robust quantitative data. 14,15 …”

mentioning

confidence: 99%

Directly Coupled High-Performance Liquid Chromatography–Accelerator Mass Spectrometry Measurement of Chemically Modified Protein and Peptides

et al. 2013

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Quantitation of low-abundance protein modifications involves significant analytical challenges, especially in biologically important applications, such as studying the role of post-translational modifications in biology and measurement of the effects of reactive drug metabolites. 14C labeling combined with accelerator mass spectrometry (AMS) provides exquisite sensitivity for such experiments. Here, we demonstrate real-time 14C quantitation of high-performance liquid chromatography (HPLC) separations by liquid sample accelerator mass spectrometry (LS-AMS). By enabling direct HPLC-AMS coupling, LS-AMS overcomes several major limitations of conventional HPLC-AMS, where individual HPLC fractions must be collected and converted to graphite before measurement. To demonstrate LS-AMS and compare the new technology to traditional solid sample AMS (SS-AMS), reduced and native bovine serum albumin (BSA) was modified by 14C-iodoacetamide, with and without glutathione present, producing adducts on the order of 1 modification in every 106 to 108 proteins. 14C incorporated into modified BSA was measured by solid carbon AMS and LS-AMS. BSA peptides were generated by tryptic digestion. Analysis of HPLC-separated peptides was performed in parallel by LS-AMS, fraction collection combined with SS-AMS, and (for peptide identification) electrospray ionization and tandem mass spectrometry (ESI-MS/MS). LS-AMS enabled 14C quantitation from ng sample sizes and was 100 times more sensitive to 14C incorporated in HPLC-separated peptides than SS-AMS, resulting in a lower limit of quantitation of 50 zmol 14C/peak. Additionally, LS-AMS turnaround times were minutes instead of days, and HPLC trace analyses required 1/ 6th the AMS instrument time required for analysis of graphite fractions by SS-AMS.

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Novel use of accelerator mass spectrometry for the quantification of low levels of systemic therapeutic recombinant protein

Cited by 19 publications

References 9 publications

Accelerator mass spectrometry of ultra-small samples with applications in the biosciences

Accelerator mass spectrometry of ultra-small samples with applications in the biosciences

Human Microdosing/Phase 0 Studies to Accelerate Drug Development

Directly Coupled High-Performance Liquid Chromatography–Accelerator Mass Spectrometry Measurement of Chemically Modified Protein and Peptides

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