The Experts below are selected from a list of 73200 Experts worldwide ranked by ideXlab platform

John Wong - One of the best experts on this subject based on the ideXlab platform.

  • immobilized metal ion Affinity Chromatography a review on its applications
    Applied Microbiology and Biotechnology, 2012
    Co-Authors: Randy Chi Fai Cheung, John Wong
    Abstract:

    After 35 years of development, immobilized metal ion Affinity Chromatography (IMAC) has evolved into a popular protein purification technique. This review starts with a discussion of its mechanism and advantages. It continues with its applications which include the purification of histidine-tagged proteins, natural metal-binding proteins, and antibodies. IMAC used in conjunction with mass spectroscopy for phosphoprotein fractionation and proteomics is also covered. Finally, this review addresses the developments, limitations, and considerations of IMAC in the biopharmaceutical industry.

James T Kadonaga - One of the best experts on this subject based on the ideXlab platform.

  • purification of sequence specific dna binding proteins by Affinity Chromatography
    Current protocols in protein science, 1993
    Co-Authors: Leslie A Kerrigan, James T Kadonaga
    Abstract:

    The Affinity Chromatography procedure described in this unit uses DNA containing specific recognition sites for the desired protein that has been covalently linked to a solid support. Preparation of a DNA Affinity resin, including cyanogen bromide (CNBr) activation of the agarose support, is described, and an alternate protocol provides a method to couple DNA to commercially available CNBr-activated Sepharose. A method for purification of crude synthetic oligonucleotides by gel electrophoresis prior to preparation of the Affinity resin is also provided. A detailed protocol for the actual Affinity Chromatography procedure is described and a support protocol allows the investigator to determine the appropriate type and quantity of nonspecific competitor DNA that should be used in the procedure and its preparation. Parameters essential to the success of an Affinity Chromatography experiment are discussed in detail in the Commentary.

Gabriella Massolini - One of the best experts on this subject based on the ideXlab platform.

  • frontal Affinity Chromatography in characterizing immobilized receptors
    Journal of Pharmaceutical and Biomedical Analysis, 2011
    Co-Authors: Enrica Calleri, Caterina Temporini, Gabriella Massolini
    Abstract:

    The state-of-the-art in frontal Affinity Chromatography (FAC) applied to receptor of pharmaceutical interest is here reported. This review will first discuss the principles of FAC for ligand characterization (K(d) determination) and for screening studies, and will examine the different strategies that have been followed for the immobilization of a broad range of receptors (cytosolic and membrane receptors). Several reported applications will then be presented demonstrating that FAC is an interesting tool enabling convenient and efficient screening in the identification of new potential ligands. Moreover new applications of FAC including dual binding site assay, receptor subtype characterization, and multi-receptor binding experiments will be underlined.

  • target based drug discovery the emerging success of frontal Affinity Chromatography coupled to mass spectrometry
    ChemMedChem, 2009
    Co-Authors: Enrica Calleri, Caterina Temporini, Gabriele Caccialanza, Gabriella Massolini
    Abstract:

    Frontal Affinity Chromatography coupled to mass spectrometry (FAC-MS) has been reported as a potential method for screening compound mixtures against immobilized target proteins. The potentiality of this analytical approach is described and illustrated with a number of examples based on targets of pharmaceutical interest.

Randy Chi Fai Cheung - One of the best experts on this subject based on the ideXlab platform.

  • immobilized metal ion Affinity Chromatography a review on its applications
    Applied Microbiology and Biotechnology, 2012
    Co-Authors: Randy Chi Fai Cheung, John Wong
    Abstract:

    After 35 years of development, immobilized metal ion Affinity Chromatography (IMAC) has evolved into a popular protein purification technique. This review starts with a discussion of its mechanism and advantages. It continues with its applications which include the purification of histidine-tagged proteins, natural metal-binding proteins, and antibodies. IMAC used in conjunction with mass spectroscopy for phosphoprotein fractionation and proteomics is also covered. Finally, this review addresses the developments, limitations, and considerations of IMAC in the biopharmaceutical industry.

Leslie A Kerrigan - One of the best experts on this subject based on the ideXlab platform.

  • purification of sequence specific dna binding proteins by Affinity Chromatography
    Current protocols in protein science, 1993
    Co-Authors: Leslie A Kerrigan, James T Kadonaga
    Abstract:

    The Affinity Chromatography procedure described in this unit uses DNA containing specific recognition sites for the desired protein that has been covalently linked to a solid support. Preparation of a DNA Affinity resin, including cyanogen bromide (CNBr) activation of the agarose support, is described, and an alternate protocol provides a method to couple DNA to commercially available CNBr-activated Sepharose. A method for purification of crude synthetic oligonucleotides by gel electrophoresis prior to preparation of the Affinity resin is also provided. A detailed protocol for the actual Affinity Chromatography procedure is described and a support protocol allows the investigator to determine the appropriate type and quantity of nonspecific competitor DNA that should be used in the procedure and its preparation. Parameters essential to the success of an Affinity Chromatography experiment are discussed in detail in the Commentary.