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Full-Text Articles in Physical Sciences and Mathematics
Development Of Affinity Microcolumns For Drug–Protein Binding Studies In Personalized Medicine: Interactions Of Sulfonylurea Drugs With In Vivo Glycated Human Serum Albumin, Jeanethe Anguizola, K. S. Joseph, Omar S. Barnaby, Ryan Matsuda, Guadalupe Alvarado, William Clarke, Ronald Cerny, David S. Hage
Development Of Affinity Microcolumns For Drug–Protein Binding Studies In Personalized Medicine: Interactions Of Sulfonylurea Drugs With In Vivo Glycated Human Serum Albumin, Jeanethe Anguizola, K. S. Joseph, Omar S. Barnaby, Ryan Matsuda, Guadalupe Alvarado, William Clarke, Ronald Cerny, David S. Hage
David Hage Publications
This report used high-performance affinity microcolumns to examine the changes in binding by sulfonylurea drugs to in vivo glycated HSA that had been isolated from individual patients with diabetes. An immunoextraction approach was developed to isolate HSA and glycated HSA from clinical samples, using only 20 μL of plasma or serum and 6–12 nmol of protein to prepare each affinity microcolumn. It was found that the affinity microcolumns could be used in either frontal analysis or zonal elution studies, which typically required only 4–8 min per run. The microcolumns had good stability and allowed data to be obtained for multiple …
Affinity Chromatography: A Review Of Clinical Applications, David S. Hage
Affinity Chromatography: A Review Of Clinical Applications, David S. Hage
David Hage Publications
Affinity chromatography is a type of liquid chromatography that makes use of biological-like interactions for the separation and specific analysis of sample components. This review describes the basic principles of affinity chromatography and examines its use in the testing of clinical samples, with an emphasis on HPLCbased methods. Some traditional applications of this approach include the use of boronate, lectin, protein A or protein G, and immunoaffinity supports for the direct quantification of solutes. Newer techniques that use antibody-based columns for on- or off-line sample extraction are examined in detail, as are methods that use affinity chromatography in combination with …
Improved Recovery Of A Radlolabeled Peptide With An Albumin-Treated Reversed-Phase Hplc Column, David S. Hage, Robert L. Taylor, Pai C. Kao
Improved Recovery Of A Radlolabeled Peptide With An Albumin-Treated Reversed-Phase Hplc Column, David S. Hage, Robert L. Taylor, Pai C. Kao
David Hage Publications
Reversed-phase high-performance liquid chromatography (RP-HPLC) is an important tool in the purification of radiolabeled peptides and proteins for immunoassay. However, for some proteins and peptides it is difficult to achieve reproducible behavior in RP-HPLC because of the low recovery of these compounds. Factors that can be varied to improve recovery include the strength or pH of the mobile phase, the chain length and spacing of groups on the reversed-phase support, and the flow rate or steepness of the elution gradient (1-5). ... In summary, we obtained better recovery and more reproducible chromatographic behavior for labeled 1-34 PTHrP with an albumin-pretreated …
Intact Parathyroid Hormone: Performance And Clinical Utility Of An Automated Assay Based On High-Performance Immunoaffinity Chromatography And Chemiluminescence Detection, David S. Hage, Bob Taylor, Pai C. Kao
Intact Parathyroid Hormone: Performance And Clinical Utility Of An Automated Assay Based On High-Performance Immunoaffinity Chromatography And Chemiluminescence Detection, David S. Hage, Bob Taylor, Pai C. Kao
David Hage Publications
The performance and clinical utility of an automated assay of intact parathyroid hormone (parathyrin, PTH) are evaluated. The method is based on the extraction of PTH from plasma by an HPLC column containing immobilized anti-(44-68 PTH) antibodies. The PTH retained is detected with a postcolumn reactor and use of anti-(1--34 PTH) chemiluminescent-labeled antibodies. The total cycle time of the assay is 6.5 mm per injection after a 1-h incubation.The lower limit of detection for PTH in a 66-pL plasma sample was 0.5 pmol/L based on peak heights and 0.2 pmol/L based on peak areas. Mean analytical recovery for PTH added …