| 1. |
- Phu, Vu Dinh, et al.
(författare)
-
Ventilator-associated respiratory infection in a resource-restricted setting: impact and etiology
- 2017
-
Ingår i: Journal of Intensive Care. - : BioMed Central (BMC). - 2052-0492. ; 5
-
Tidskriftsartikel (refereegranskat)abstract
- Ventilator-associated respiratory infection (VARI) is a significant problem in resource-restricted intensive care units (ICUs), but differences in casemix and etiology means VARI in resource-restricted ICUs may be different from that found in resource-rich units. Data from these settings are vital to plan preventative interventions and assess their cost-effectiveness, but few are available.
|
|
| 2. |
- Hoang-Minh, Thao, et al.
(författare)
-
Use of TEM-EDX for structural formula identification of clay minerals : a case study of Di Linh bentonite, Vietnam
- 2019
-
Ingår i: Journal of applied crystallography. - : International Union of Crystallography (IUCr). - 0021-8898 .- 1600-5767. ; 52:1, s. 133-147
-
Tidskriftsartikel (refereegranskat)abstract
- Transmission electron microscopy linked with energy-dispersive X-ray spectroscopy (TEM-EDX) was applied to characterize mineralogical signals ofweathering processes in the Di Linh bentonite deposit (Vietnam) and to visualize the effects of Na activation on the smectitic phases. Modelling of X ray diffraction patterns (oriented mount) was applied in order to refine the computed structural formula. X-ray diffraction, X-ray fluorescence and Fouriertransform infrared spectroscopy (FT-IR) methods were also applied to verify the TEM-EDX results. An Excel-based routine has been developed in this research to allow fast computation of structural formulae and classification of the investigated clay particles. This routine supports the acquirement of 100 300 TEM-EDX analyses as a representative set of individual particles for each sample. The Excel-based routine involves end members of different clay mineral groups and interstratifications with two or three members (e.g. illite smectite interstratifications – IS-ml; dioctahedral vermiculite–smectite interstratifications – diVS-ml; and kaolinite–montmorillonite–dioctahedral vermiculite interstratifications – KSV-ml). The routine is now freely available. According to the identification procedure, the <2 mm fraction of the Di Linh bentonite (Vietnam) is composed mainly of K- and charge-deficient illite smectite interstratifications (or diVS-ml): montmorillonite-rich randomly ordered (R0) type and illite-rich regularly ordered (R1) type. Additionally, Fe-poor KSV-ml was identified.Industrial Na activation of the Di Linh bentonite resulted in an increase of theR1 diVS-ml portion and dissolution of a large part of the smectite-rich phases.The TEM-EDX approach also gave analytical proof of a sedimentary processfor Di Linh smectite. The parent muscovite was altered in two different environments: (i) K-leaching and layer-wise alteration into kaolinite (weathering), and (ii) further edge-controlled alteration of mica into lath-like montmorillonite particles associated with a dissolution of kaolinite layers from the former kaolinite–mica intergrowths by heat impact (basalt flow).
|
|
| 3. |
- Duong-Thi, Minh-Dao, et al.
(författare)
-
Comparison of weak affinity chromatography and surface plasmon resonance in determining affinity of small molecules
- 2014
-
Ingår i: Analytical Biochemistry. - : Elsevier BV. - 0003-2697 .- 1096-0309. ; 461, s. 57-59
-
Tidskriftsartikel (refereegranskat)abstract
- In this study, we compared affinity data from surface plasmon resonance (SPR) and weak affinity chromatography (WAC), two established techniques for determination of weak affinity (mM-mu M) small molecule-protein interactions. In the current comparison, thrombin was used as target protein. In WAC the affinity constant (K-D) was determined from retention times, and in SPR it was determined by Langmuir isotherm fitting of steady-state responses. Results indicate a strong correlation between the two methods (R-2 = 0.995, P < 0.0001). (C) 2014 Elsevier Inc. All rights reserved.
|
|
| 4. |
- Duong-Thi, Minh-Dao, et al.
(författare)
-
High-Throughput Fragment Screening by Affinity LC-MS
- 2013
-
Ingår i: Journal of Biomolecular Screening. - : Elsevier BV. - 1087-0571 .- 1552-454X. ; 18:2, s. 160-171
-
Tidskriftsartikel (refereegranskat)abstract
- Fragment screening, an emerging approach for hit finding in drug discovery, has recently been proven effective by its first approved drug, vemurafenib, for cancer treatment. Techniques such as nuclear magnetic resonance, surface plasmon resonance, and isothemal titration calorimetry, with their own pros and cons, have been employed for screening fragment libraries. As an alternative approach, screening based on high-performance liquid chromatography separation has been developed. In this work, we present weak affinity LC/MS as a method to screen fragments under high-throughput conditions. Affinity-based capillary columns with immobilized thrombin were used to screen a collection of 590 compounds from a fragment library. The collection was divided into 11 mixtures (each containing 35 to 65 fragments) and screened by MS detection. The primary screening was performed in < 4 h (corresponding to > 3500 fragments per day). Thirty hits were defined, which subsequently entered a secondary screening using an active site-blocked thrombin column for confirmation of specificity. One hit showed selective binding to thrombin with an estimated dissociation constant (K-D) in the 0.1 mM range. This study shows that affinity LC/MS is characterized by high throughput, ease of operation, and low consumption of target and fragments, and therefore it promises to be a valuable method for fragment screening.
|
|
| 5. |
- Duong-Thi, Minh-Dao
(författare)
-
Introducing weak affinity chromatography to drug discovery with focus on fragment screening
- 2013
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Fragment-based drug discovery is an emerging process that has gained popularity in recent years. The process starts from small molecules called fragments. One major step in fragment-based drug discovery is fragment screening, which is a strategy to screen libraries of small molecules to find hits. The strategy in theory is more efficient than traditional high-throughput screening that works with larger molecules. As fragments intrinsically possess weak affinity to a target, detection techniques of high sensitivity to affinity are required for fragment screening. Furthermore, the use of different screening methods is necessary to improve the likelihood of success in finding suitable fragments. Since no single method can work for all types of screening, there is a demand for new techniques. The aim of this thesis is to introduce weak affinity chromatography (WAC) as a novel technique for fragment screening.WAC is, as the name suggests, an affinity-based liquid chromatographic technique that separates compounds based on their different weak affinities to an immobilized target. The higher affinity a compound has towards the target, the longer it remains in the separation unit, and this will be expressed as a longer retention time. The affinity measure and ranking of affinity can be achieved by processing the obtained retention times of analyzed compounds.In this thesis, WAC is studied for fragment screening on two platforms. The first system comprised a 24-channel affinity cartridge that works in cooperation with an eight-needle autosampler and 24 parallel UV detector units. The second system was a standard analytical LC-MS platform that is connected to an affinity column, generally called WAC-MS or affinity LC-MS. The evaluation criteria in studying WAC for fragment screening using these platforms were throughput, affinity determination and ranking, specificity, operational platform characteristics and consumption of target protein and sample. The model target proteins were bovine serum albumin for the first platform, thrombin and trypsin for the latter. Screened fragments were either small molecule drugs, a thrombin-directed collection of compounds, or a general-purpose fragment library. To evaluate WAC for early stages of fragment elaboration, diastereomeric mixtures from a thrombin-directed synthesis project were screened.Although both analytical platforms can be used for fragment screening, WAC-MS shows more useful features due to easy access to the screening platform, higher throughput and ability to analyze mixtures. Affinity data from WAC are in good correlation with IC50 values from enzyme assay experiments. The possibility to distinguish specific from non- specific interactions plays an important role in the interpretation of WAC results. In this thesis, this was achieved by inhibiting the active site of the target protein to measure off-site interactions. WAC proves to be a sensitive, robust, moderate in cost and easy to access technique for fragment screening, and can also be useful in the early stages of fragment evolution.In conclusion, this thesis has demonstrated the proof of principle of using WAC as a new tool to monitor affinity and to select hits in fragment-based drug discovery. This thesis has indicated the primary possibilities, advantages as well as the limitations of WAC in fragment screening procedures. In the future, WAC should be evaluated on other targets and fragment libraries in order to realize more fully the potential of the technology.
|
|
| 6. |
- Duong-Thi, Minh-Dao, et al.
(författare)
-
Lipodisks integrated with weak affinity chromatography enable fragment screening of integral membrane proteins
- 2016
-
Ingår i: The Analyst. - : Royal Society of Chemistry (RSC). - 0003-2654 .- 1364-5528. ; 141:3, s. 981-988
-
Tidskriftsartikel (refereegranskat)abstract
- Membrane proteins constitute the largest class of drug targets but they present many challenges in drug discovery. Importantly, the discovery of potential drug candidates is hampered by the limited availability of efficient methods for screening drug-protein interactions. In this work we present a novel strategy for rapid identification of molecules capable of binding to a selected membrane protein. An integral membrane protein (human aquaporin-1) was incorporated into planar lipid bilayer disks (lipodisks), which were subsequently covalently coupled to porous derivatized silica and packed into HPLC columns. The obtained affinity columns were used in a typical protocol for fragment screening by weak affinity chromatography (WAC), in which one hit was identified out of a 200 compound collection. The lipodisk-based strategy, which ensures a stable and native-like lipid environment for the protein, is expected to work also with other membrane proteins and screening procedures.
|
|
| 7. |
- Duong-Thi, Minh-Dao, et al.
(författare)
-
Lipodisks integrated with weak affinity chromatography enable fragment screening of integral membrane proteins
- 2016
-
Ingår i: The Analyst. - 0003-2654 .- 1364-5528. ; 141:3, s. 981-988
-
Tidskriftsartikel (refereegranskat)abstract
- Membrane proteins constitute the largest class of drug targets but they present many challenges in drug discovery. Importantly, the discovery of potential drug candidates is hampered by the limited availability of efficient methods for screening drug-protein interactions. In this work we present a novel strategy for rapid identification of molecules capable of binding to a selected membrane protein. An integral membrane protein (human aquaporin-1) was incorporated into planar lipid bilayer disks (lipodisks), which were subsequently covalently coupled to porous derivatized silica and packed into HPLC columns. The obtained affinity columns were used in a typical protocol for fragment screening by weak affinity chromatography (WAC), in which one hit was identified out of a 200 compound collection. The lipodisk-based strategy, which ensures a stable and native-like lipid environment for the protein, is expected to work also with other membrane proteins and screening procedures.
|
|
| 8. |
- Duong-Thi, Minh-Dao, et al.
(författare)
-
Weak affinity chromatography as a new approach for fragment screening in drug discovery
- 2011
-
Ingår i: Analytical Biochemistry. - : Elsevier. - 0003-2697 .- 1096-0309. ; 414:1, s. 138-146
-
Tidskriftsartikel (refereegranskat)abstract
- Fragment-based drug design (FBDD) is currently being implemented in drug discovery, creating a demand for developing efficient techniques for fragment screening. Due to the intrinsic weak or transient binding of fragments (mM–uM in dissociation constant (KD)) to targets, methods must be sensitive enough to accurately detect and quantify an interaction. This study presents weak affinity chromatography (WAC) as an alternative tool for screening of small fragments. The technology was demonstrated by screening of a selected 23 compound fragment collection of documented binders, mostly amidines, using trypsin and thrombin as model target protease proteins. WAC was proven to be a sensitive, robust, and reproducible technique that also provides information about affinity of a fragment in the range of 1 mM–10uM. Furthermore, it has potential for high throughput as was evidenced by analyzing mixtures in the range of 10 substances by WAC–MS. The accessibility and flexibility of the technology were shown as fragment screening can be performed on standard HPLC equipment. The technology can further be miniaturized and adapted to the requirements of affinity ranges of the fragment library. All these features of WAC make it a potential method in drug discovery for fragment screening.
|
|
| 9. |
- Duong-Thi, Minh-Dao, et al.
(författare)
-
Weak Affinity Chromatography for Evaluation of Stereoisomers in Early Drug Discovery
- 2013
-
Ingår i: Journal of Biomolecular Screening. - : Sage Publications. - 1087-0571 .- 1552-454X. ; 18:6, s. 748-755
-
Tidskriftsartikel (refereegranskat)abstract
- In early drug discovery (e.g. in fragment screening), recognition of stereoisomeric structures is valuable and guides medicinal chemists to focus only on useful configurations. In this work, we concurrently screened mixtures of stereoisomers and estimated their affinities to a protein target (thrombin) using weak affinity chromatography-mass spectrometry (WAC-MS). Affinity determinations by WAC showed that minor changes in stereoisomeric configuration could have major impact on affinity. The ability of WAC-MS to provide instant information about stereoselectivity and binding affinities directly from analyte mixtures is a great advantage in fragment library screening and drug lead development.
|
|
| 10. |
|
|
| 11. |
- Ohlson, Sten, et al.
(författare)
-
Toward high-throughput drug screening on a chip-based parallell affinity separation platform
- 2010
-
Ingår i: Journal of Separation Science. - : Wiley. - 1615-9306 .- 1615-9314. ; 33:17-18, s. 2575-2581
-
Tidskriftsartikel (refereegranskat)abstract
- High-throughput screening of compound libraries, including the study of fragments, has become one of the cornerstones in modern drug discovery research. During this process hits are defined that may be developed into valuable leads and eventually into possible drug candidates. In this paper, we have demonstrated that parallel zonal weak affinity chromatography in microcolumns on a chip offers a possible screening format for weakly binding ligands toward a protein target. We used albumin as a model system because this transport protein is well established as a binder (both weak and strong) for drug substances. Bovine serum albumin was immobilized on microparticulate diolsilica particles and then packed into a 24-channel cartridge, which served as the separation platform. Analysis of the obtained chromatograms yielded information about affinity even in the millimolar range. Employing this approach, thousands of substances can be screened in just a day. We feel confident that zonal affinity chromatography will provide a useful technology in the future for performing high-throughput screening.
|
|
