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Affinity Chromatography

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Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
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Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
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Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
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The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
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Principles Of Column Chromatography01:13

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The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...
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Nanopillar-Assisted SERS Chromatography.

Onur Durucan1, Kaiyu Wu1, Marlitt Viehrig1

  • 1DNRF and Villum Fonden Center for Intelligent Drug Delivery and Sensing Using Microcontainers and Nanomechanics, IDUN, Department of Micro- and Nanotechnology , Technical University of Denmark , Kgs. Lyngby 2800 , Denmark.

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This study introduces nanopillar-assisted SERS chromatography (NPC-SERS), a new method simplifying complex sample analysis. NPC-SERS enables rapid, quantitative detection of molecules in biofluids like urine without extensive pretreatment.

Keywords:
full automationmulticomponent analysisnanopillar-assisted chromatographyquantitative SERSreal-life samples

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Area of Science:

  • Analytical Chemistry
  • Spectroscopy
  • Microfluidics

Background:

  • Surface-enhanced Raman spectroscopy (SERS) faces challenges with complex samples requiring extensive purification.
  • Current methods for analyzing biofluids are often time-consuming and costly.

Purpose of the Study:

  • To develop a novel method for simultaneous quantitation and analysis of target molecules in complex fluids.
  • To overcome limitations of traditional SERS by simplifying sample handling and enhancing applicability.

Main Methods:

  • Developed nanopillar-assisted SERS chromatography (NPC-SERS) combining gold-coated silicon nanopillars (AuNP) with a centrifugal microfluidic platform.
  • Utilized the 'wicking effect' for efficient wetting of the stationary phase, minimizing analyte dilution.
  • Demonstrated spatial separation of molecules on the AuNP substrate for automated sample manipulation.

Main Results:

  • Successfully separated paracetamol (PAR) from major human urine components (urea, uric acid, creatinine).
  • Achieved quantitative detection of PAR with an ultrawide linear dynamic range (0-500 ppm) by analyzing spreading profiles.
  • Validated the NPC-SERS technique for analyzing complex biofluid samples.

Conclusions:

  • NPC-SERS offers a streamlined approach for analyzing complex biological samples, transforming SERS into a broadly applicable sensing technique.
  • Facilitates rapid, quantitative detection of analytes in diverse biofluids including urine, saliva, and blood.
  • Reduces the need for costly and time-consuming sample pretreatment steps.