Principal Component Analysis of 1D 1H Diffusion Edited NMR Spectra of Protein Therapeutics.

The one-dimensional (1D) diffusion edited proton NMR method, Protein Fingerprint by Lineshape Enhancement (PROFILE) has been demonstrated to be suitable for higher order structure (HOS) characterization of protein therapeutics including monoclonal antibodies. Recent reports in the literature have demonstrated its advantages for HOS characterization over traditional methods such as circular dichroism and Fourier-transform infrared spectroscopy. Previously, we have demonstrated that the PROFILE method is complementary with high resolution 2D methyl correlated NMR methods and how both may be deployed as a multi-modal platform to further the utility of NMR for HOS characterization. A major limitation of the PROFILE method remains its need for high signal to noise data due to its reliance on convolution difference processing and linear correlation metrics to assess spectral similarity. Here we present an alternative method for analyzing 1D diffusion edited spectra, which overcomes this limitation by using nonlinear iterative partial least squares (NIPALS) principal component analysis, and which we dub PROtein Fingerprint Observed Using NIPALS Decomposition (PROFOUND). We demonstrate that results from the PROFOUND method are robust with respect to instrument, operator and in the presence of high experimental noise and how it may be employed to provide quantitative assessment of spectral similarity.

Comparative Analysis of One-Dimensional Protein Fingerprint by Line Shape Enhancement and Two-Dimensional 1H,13C Methyl NMR Methods for Characterization of the Higher Order Structure of IgG1 Monoclonal Antibodies.

The use of NMR spectroscopy has emerged as a premier tool to characterize the higher order structure of protein therapeutics and in particular IgG1 monoclonal antibodies (mAbs). Due to their large size, traditional 1H-15N correlation experiments have proven exceedingly difficult to implement on mAbs, and a number of alternative techniques have been proposed, including the one-dimensional (1D) 1H protein fingerprint by line shape enhancement (PROFILE) method and the two-dimensional (2D) 1H-13C methyl correlation-based approach. Both 1D and 2D approaches have relative strengths and weaknesses, related to the inherent sensitivity and resolution of the respective methods. To further increase the utility of NMR to the biopharmaceutical community, harmonized criteria for decision making in employing 1D and 2D approaches for mAb characterization are warranted. To this end, we have conducted an interlaboratory comparative study of the 1D PROFILE and 2D methyl methods on several mAbs samples to determine the degree to which each method is suited to detect spectral difference between the samples and the degree to which results from each correlate with one another. Results from the study demonstrate both methods provide statistical data highly comparable to one another and that each method is capable of complementing the limitations commonly associated with the other, thus providing a better overall picture of higher order structure.

Narrowing the filter-cavity bandwidth in gravitational-wave detectors via optomechanical interaction.

We propose using optomechanical interaction to narrow the bandwidth of filter cavities for achieving frequency-dependent squeezing in advanced gravitational-wave detectors, inspired by the idea of optomechanically induced transparency. This can allow us to achieve a cavity bandwidth on the order of 100 Hz using small-scale cavities. Additionally, in contrast to a passive Fabry-Pérot cavity, the resulting cavity bandwidth can be dynamically tuned, which is useful for adaptively optimizing the detector sensitivity when switching amongst different operational modes. The experimental challenge for its implementation is a stringent requirement for very low thermal noise of the mechanical oscillator, which would need a superb mechanical quality factor and a very low temperature. We consider one possible setup to relieve this requirement by using optical dilution to enhance the mechanical quality factor.

Multicolor cavity metrology.

Long-baseline laser interferometers used for gravitational-wave detection have proven to be very complicated to control. In order to have sufficient sensitivity to astrophysical gravitational waves, a set of multiple coupled optical cavities comprising the interferometer must be brought into resonance with the laser field. A set of multi-input, multi-output servos then lock these cavities into place via feedback control. This procedure, known as lock acquisition, has proven to be a vexing problem and has reduced greatly the reliability and duty factor of the past generation of laser interferometers. In this article, we describe a technique for bringing the interferometer from an uncontrolled state into resonance by using harmonically related external fields to provide a deterministic hierarchical control. This technique reduces the effect of the external seismic disturbances by 4 orders of magnitude and promises to greatly enhance the stability and reliability of the current generation of gravitational-wave detectors. The possibility for using multicolor techniques to overcome current quantum and thermal noise limits is also discussed.

Presence and metabolism of endogenous androgenic-anabolic steroid hormones in meat-producing animals: a review.

The presence and metabolism of endogenous steroid hormones in meat-producing animals has been the subject of much research over the past 40 years. While significant data are available, no comprehensive review has yet been performed. Species considered in this review are bovine, porcine, ovine, equine, caprine and cervine, while steroid hormones include the androgenic-anabolic steroids testosterone, nandrolone and boldenone, as well as their precursors and metabolites. Information on endogenous steroid hormone concentrations is primarily useful in two ways: (1) in relation to pathological versus 'normal' physiology and (2) in relation to the detection of the illegal abuse of these hormones in residue surveillance programmes. Since the major focus of this review is on the detection of steroids abuse in animal production, the information gathered to date is used to guide future research. A major deficiency in much of the existing published literature is the lack of standardization and formal validation of experimental approach. Key articles are cited that highlight the huge variation in reported steroid concentrations that can result when samples are analysed by different laboratories under different conditions. These deficiencies are in most cases so fundamental that it is difficult to make reliable comparisons between data sets and hence it is currently impossible to recommend definitive detection strategies. Standardization of the experimental approach would need to involve common experimental protocols and collaboratively validated analytical methods. In particular, standardization would need to cover everything from the demographic of the animal population studied, the method of sample collection and storage (especially the need to sample live versus slaughter sampling since the two methods of surveillance have very different requirements, particularly temporally), sample preparation technique (including mode of extraction, hydrolysis and derivatization), the end-point analytical detection technique, validation protocols, and the statistical methods applied to the resulting data. Although efforts are already underway (at HFL and LABERCA) to produce more definitive data and promote communication among the scientific community on this issue, the convening of a formal European Union working party is recommended.

Monitoring of pesticides and veterinary drugs in Australian cattle: verification of the residue control system.

The Australian National Residue Survey in partnership with the Australian Quarantine and Inspection Service and industry routinely monitors beef to ensure that regulatory requirements for residues are being met. In the 10-year period from 1997 to 2006 a total of 128,902 samples were subjected to residue testing covering veterinary drugs and veterinary and agricultural pesticides in a random monitoring program. Residues of agricultural and veterinary chemicals exceeded the Australian standard in 13 and 7 samples, respectively. Greater than 99% of samples did not contain detectable residues above the Australian standard while greater than 90% of samples did not contain any residues of compounds included in the survey. The results of the surveys provide verification that the Australian residue control system is appropriate and that the requirements of countries importing Australian meat products can be satisfied. The surveys have also demonstrated that industry based programs and state based regulatory controls over sites contaminated with organochlorine residues are effective in managing residues of these chemicals in cattle.

Estimation of measurement uncertainty in pesticide residue analysis.

Proficiency test results from 5 countries involving 61 separate interlaboratory proficiency tests for pesticide residues were examined in this study. A total of 24 different matrixes and 869 relative standard deviations of the mean (or median) pesticide residue concentration were statistically evaluated in relation to the Horwitz function. The aim was to determine whether or not the concentration-dependent relationship described by Horwitz would hold for the much narrower range of chemicals and concentrations covered in routine pesticide residue analysis. Although for fatty (animal-derived) matrixes the variability increased as the concentration decreased in line with the Horwitz equation, the between-laboratories relative standard deviations for nonfatty matrixes (fruit, vegetables, and grain) remained at 25% over the entire concentration range of 1 microg/kg to 10 mg/kg for the pesticides studied. Given these findings, the Horwitz equation remains valid for calculating uncertainties involving pesticide residues in fatty matrixes. However, for pesticide residue analyses involving nonfatty matrixes, a constant relative standard deviation of 25% is more appropriate for calculating uncertainties, particularly when a reported result is assessed against a regulatory limit.

Ion-chromatographic determination of chloride and fluoride in electrolyte from the halogen tin-plating process.

A simple and rapid procedure is proposed for the determination of chloride and free fluoride in tin electroplating fluid. Suppressor-column ion-chromatography is used after oxidation of hexafluorostannate(II) to hexafluorostannate(IV) with hydrogen peroxide. Concurrent determination of tin(II) and total tin then allows calculation of the concentrations of fluoride, hexafluorostannate(II) and hexafluorostannate(IV).