Abstract:A parallel nanoliter microfluidic analysis system based on capillary action, centrifugal force, and hydrophobic barriers is described. The precision of 112 parallel volume definition operations is determined to 0.75% CV at 200 nL using the individual sample introduction structure. For 20 nL, the actual measurement error is the dominating factor, with a combined error of 1.9% CV. Individual dispensing as well as dispensing through a common distribution channel is described. The volume definition precision for t… Show more
“…IgG1 was captured from a crude cell supernatant using protein A beads and the N-glycans were released with PNGase F. Glycans were purified with a graphitized carbon black column, and both this stage and crystallization for MALDI-TOF analysis were performed on the CD (Andersson et al, 2007;Thuy et al, 2010). Tep et al (2012b) have developed a method for looking at glycomic changes that occur in CHO cells other than those associated with the expressed biotherapeutic.…”
“…IgG1 was captured from a crude cell supernatant using protein A beads and the N-glycans were released with PNGase F. Glycans were purified with a graphitized carbon black column, and both this stage and crystallization for MALDI-TOF analysis were performed on the CD (Andersson et al, 2007;Thuy et al, 2010). Tep et al (2012b) have developed a method for looking at glycomic changes that occur in CHO cells other than those associated with the expressed biotherapeutic.…”
“…11) is an open, parallel nanoliter microfluidic analysis system [18] which allows the development of new assays if suitable biotinylated antigens are available. The streptavidin-coated beads are activated with biotinylated capture reagents as the first step in the immunoassay.…”
Section: Technical Trends In Immunoassaysmentioning
Laboratory measurements have become an essential part of medical diagnostics. In pediatric endocrinology, measurements of hormones and related biomarkers like growth factors are practically indispensable. They initially allow the evaluation of the pathological stage and differential diagnosis of a patient. In the following, they are used to monitor and predict a therapeutic success and outcome. Thus, measurements of hormones and growth factors must be valid and reproducible in both technical and medical terms. The laboratory must warrant the technical validity and reproducibility of the hormone measurements through standardized laboratory procedures and adequate quality standards and quality management. It is, however, the responsibility of the treating physician, first to choose the right laboratory parameters to be examined, and second to ensure adequate test circumstances including the time of sampling, influence of nutrition and cyclic factors, co-medication and shipment of the sample. The physician/pediatrician is responsible for interpretation of the laboratory results and the use of this medical information in patient care.During the last two decades, laboratory diagnostics in endocrinology and in general have undergone a technological revolution in several ways. The major trends are the following.
Multiplex Techniques and MiniaturizationMultiplex techniques represent a current megatrend in laboratory diagnostics with the aim of saving time and resources upon the release of the diagnostic result, and
“…In contrast to pressure-driven microfluidic approaches, centrifugal microfluidic systems can be filled with regular pipettes and intrinsically benefit from low dead volumes since no liquid is lost in tubings. 19 This simple "world-to-chip" interface and low dead volumes have led to the adoption of centrifugal microfluidic platforms in diverse diagnostic 20,21 and analytical 22 applications. Especially noteworthy are centrifugal microfluidic systems for protein crystallography.…”
We present a centrifugal microfluidic LabDisk for protein structure analysis via small-angle X-ray scattering (SAXS) on synchrotron beamlines. One LabDisk prepares 120 different measurement conditions, grouped into six dilution matrices. Each dilution matrix: (1) features automatic generation of 20 different measurement conditions from three input liquids and (2) requires only 2.5 μl of protein solution, which corresponds to a tenfold reduction in sample volume in comparison to the state of the art. Total hands on time for preparation of 120 different measurement conditions is less than 5 min. Read-out is performed on disk within the synchrotron beamline P12 at EMBL Hamburg (PETRA III, DESY). We demonstrate: (1) aliquoting of 40 nl aliquots for five different liquids typically used in SAXS and (2) confirm fluidic performance of aliquoting, merging, mixing and read-out from SAXS experiments (2.7-4.4% CV of protein concentration).We apply the LabDisk for SAXS for basic analysis methods, such as measurement of the radius of gyration, and advanced analysis methods, such as the ab initio calculation of 3D models. The suitability of the LabDisk for SAXS for protein structure analysis under different environmental conditions is demonstrated for glucose isomerase under varying protein and NaCl concentrations. We show that the apparent radius of gyration of the negatively charged glucose isomerase decreases with increasing protein concentration at low salt concentration. At high salt concentration the radius of gyration (R g ) does not change with protein concentrations. Such experiments can be performed by a non-expert, since the LabDisk for SAXS does not require attachment of tubings or pumps and can be filled with regular pipettes. The new platform has the potential to introduce routine high-throughput SAXS screening of protein structures with minimal input volumes to the regular operation of synchrotron beamlines.
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