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Full-Text Articles in Life Sciences

Analysis Of Biomolecular Interactions Using Affinity Microcolumns: A Review, Xiwei Zheng, Zhao Li, Sandya Beeram, Maria Podariu, Ryan Matsuda, Erika L. Pfaunmiller, Christopher J. White Ii, Natasha Carter, David S. Hage Jan 2014

Analysis Of Biomolecular Interactions Using Affinity Microcolumns: A Review, Xiwei Zheng, Zhao Li, Sandya Beeram, Maria Podariu, Ryan Matsuda, Erika L. Pfaunmiller, Christopher J. White Ii, Natasha Carter, David S. Hage

David Hage Publications

Affinity chromatography has become an important tool for characterizing biomolecular interactions. The use of affinity microcolumns, which contain immobilized binding agents and have volumes in the mid-to-low microliter range, has received particular attention in recent years. Potential advantages of affinity microcolumns include the many analysis and detection formats that can be used with these columns, as well as the need for only small amounts of supports and immobilized binding agents. This review examines how affinity microcolumns have been used to examine biomolecular interactions. Both capillary-based microcolumns and short microcolumns are considered. The use of affinity microcolumns with zonal elution and …


Affinity Monolith Chromatography: A Review Of Principles And Recent Analytical Applications, Erika L. Pfaunmiller, Marie Laura Paulemond, Courtney M. Dupper, David S. Hage Mar 2013

Affinity Monolith Chromatography: A Review Of Principles And Recent Analytical Applications, Erika L. Pfaunmiller, Marie Laura Paulemond, Courtney M. Dupper, David S. Hage

David Hage Publications

Affinity monolith chromatography (AMC) is a type of liquid chromatography that uses a monolithic support and a biologically-related binding agent as a stationary phase. AMC is a powerful method for the selective separation, analysis or studies of specific target compounds in a sample. This review discusses the basic principles of AMC and recent developments or applications of this method, with particular emphasis being given to work that has appeared in the last five years. Various materials that have been used to prepare columns for AMC are examined, including organic monoliths, silica monoliths, agarose monoliths and cryogels. These supports have been …


Studies Of Metabolite–Protein Interactions: A Review, Ryan Matsuda, Cong Bi, Jeanethe Anguizola, Matthew Sobansky, Elliott Rodriguez, John Vargas Badilla, Xiwei Zheng, Benjamin Hage, David S. Hage Jan 2013

Studies Of Metabolite–Protein Interactions: A Review, Ryan Matsuda, Cong Bi, Jeanethe Anguizola, Matthew Sobansky, Elliott Rodriguez, John Vargas Badilla, Xiwei Zheng, Benjamin Hage, David S. Hage

David Hage Publications

The study of metabolomics can provide valuable information about biochemical pathways and processes at the molecular level. There have been many reports that have examined the structure, identity and concentrations of metabolites in biological systems. However, the binding of metabolites with proteins is also of growing interest. This review examines past reports that have looked at the binding of various types of metabolites with proteins. An overview of the techniques that have been used to characterize and study metabolite–protein binding is first provided. This is followed by examples of studies that have investigated the binding of hormones, fatty acids, drugs …


Affinity Chromatography: A Review Of Clinical Applications, David S. Hage Jan 1999

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 Jan 1992

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 Jan 1992

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 …


Use Of Affinity Chromatography In Developing Acridinium Ester-Labeled Antibodies For An Immunometric Assay Of Parathyrin, David S. Hage, Bob Taylor, Pat Schryver, Pai C. Kao Jan 1991

Use Of Affinity Chromatography In Developing Acridinium Ester-Labeled Antibodies For An Immunometric Assay Of Parathyrin, David S. Hage, Bob Taylor, Pat Schryver, Pai C. Kao

David Hage Publications

In developing an immunometric assay of intact parathyrin (parathyroid hormone, PTH), we found that affinity chromatography is a useful tool in purifying and optimizing the labeling conditions for acridinium ester-labeled antibodies. ... In summary, affinity chromatography was found to be useful in the purification of acridinium ester-labeled antibodies, particularly for removing denatured antibodies from the prepared label and for monitoring the amount of active labeled antibodies produced.