Immobilized iminodiacetic acid (IDA)-type Cu2+-chelating membrane affinity chromatography for purification of bovine liver catalase

Over the past couple of years, proteomics pattern analysis has emerged as an effective method for the early diagnosis of diseases such as ovarian, breast, or prostate cancer, without identification of single biomarkers. MALDI-TOF MS, for example, offers a simple approach for fast and reliable protein profiling, especially by using carrier materials with various physical and chemical properties, in combination with a MALDI matrix. This approach is referred to as materialenhanced laser desorption/ionization (MELDI). In this paper, we report the development and application of derivatized carrier materials [cellulose, silica, poly(glycidyl methacrylate/divinylbenzene) (GMA/DVB) particles, and diamond powder] for fast and direct MALDI-TOF MS protein profiling. The applicability of MELDI for rapid protein profiling was evaluated with human serum samples. These carriers, having various hydrophobicities, resulted in characteristic mass fingerprints, even if all materials were derivatized with iminodiacetic acid (IDA) to yield an immobilized metal affinity chromatography (IMAC) functionality. Our study demonstrates that analyzing complex biological samples, such as human serum, by employing different MELDI carrier materials yielded type-and size-dependent performance variation. T he quest to diagnose and identify diseases at early stages depends mainly on the accurate determination of the marker entities in the biological fluids. These marker entities are normally very low concentrated [1], and highly sensitive methods are required to bind them effectively from the biosamples and to measure them with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) [2]. MALDI mass spectrometry, in recent times, has proven to be an authentic technique to analyze peptides and proteins [3,4]. These biomolecules out of the complex biofluids like serum are difficult to ionize and thus, bring into time of flight (TOF) because of the complexity of the sample and the suppressing nonvolatile salts [5]. The aforementioned problems are tackled by another technique introduced years ago, namely, surface-enhanced laser desorption/ionization time of flight mass spectrometry (SELDI-TOF MS), for the detection of several peptides and proteins from complex biological specimens [6 -8]. This affinity-based method utilizes selective adsorption of proteins on various chromatographic surfaces to reduce sample complexity [9,10]. Spectra generated by SELDI-TOF MS usually show limited peak and binding capacity due to the smaller surface area of the plane protein chip [11]. Limitations of SELDI are also associated with the lower mass resolution of the applied MS instrument that

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“…Cellulose was treated with glycidyl methacylate (Figure 1a) at elevated temperature in the presence of a redox catalyst as described in [15,16].…”

Section: Particle Preparation and Derivatizationmentioning

confidence: 99%

Material-enhanced laser desorption/ionization (MELDI)—A new protein profiling tool utilizing specific carrier materials for time of flight mass spectrometric analysis

Feuerstein

Najam‐ul‐Haq

Rainer

et al. 2006

J. Am. Soc. Mass Spectrom.

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“…The method has found widespread application in the purification of recombinant histidine‐tagged and native pharmaceutical proteins, most commonly using Cu(II)‐ and Ni(II)‐charged iminodiacetic acid (IDA) and nitrilotriacetic acid (NTA) chromatographic adsorbents. Metal chelate ligands have also been immobilized on foams (3), membranes (4), and biosensor chips (5) and in electrophoresis gels (6); they have also been used as affinity precipitation agents (7).…”

Section: Introductionmentioning

confidence: 99%

Nucleic Acid Separations Utilizing Immobilized Metal Affinity Chromatography

et al. 2003

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Immobilized metal affinity chromatography (IMAC) is widely used for protein purification, e.g., in the isolation of proteins bearing the well-known hexahistidine affinity tag. We report that IMAC matrixes can also adsorb single-stranded nucleic acids through metal ion interactions with aromatic base nitrogens and propose that metal affinity technologies may find widespread application in nucleic acid technology. Oligonucleotide duplexes, plasmid, and genomic DNA show low IMAC binding affinity, while RNA and single-stranded oligonucleotides bind strongly to matrixes such as Cu(II) iminodiacetic acid (IDA) agarose. The affinity of yeast RNA for IDA-chelated metal ions decreases in the following order: Cu(II), Ni(II), Zn(II), and Co(II). Adsorption isotherms for 20-mer oligonucleotide homopolymers show that purines are strongly favored over pyrimidines and that double-stranded duplexes are not bound. IMAC columns have been used to purify plasmid DNA from E. coli alkaline lysates, to purify a ribozyme, to remove primers and imperfect products from PCR reactions, and to separate 20-mer oligonucleotide duplexes containing centered single-base mismatches. Potential further applications include SNP scoring, hybridization assays, and the isolation of polyadenylated messenger RNA.

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“…1 IMAC separation exploits the affinity between the immobilized metal ions and electron-donating groups on the protein surface. 2 IMAC has many advantages over typical methods of affinity chromatography: different metal ions can be immobilized on the same chelating medium after stripping off the column and can be easily removed for regeneration by a stronger chelator such as EDTA, 3 and various carboxymethylated amines [4][5][6] can be used as chelating ligands for immobilization of metal ions. The optimal conditions for separation of target proteins can be obtained by proper choice of suitable metal ions as well as chelating ligand in order to keep biological activity of the protein after elution from an IMAC column.…”

Section: Introductionmentioning

confidence: 99%

Preparation of Immobilized Metal Affinity Chromatographic Packings by Immobilization of Carboxymethylated Asparate (CM‐Asp) Based on Monodisperse Hydrophilic Non‐porous Beads and Their Application

Gong

Wang

2010

Chin. J. Chem.

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The hydrophilic immobilized metal affinity chromatographic packing was prepared by immobilization of carboxymethylated asparate (CM-Asp) as chelating ligand and Ni 2＋ as center ion on the base of monodispersed, 3.0 µm non-porous monodisperse poly(glycidylmethacrylate-co-ethylenedimethacrylate) (P GMA/EDMA ) particles. The retention behavior of proteins and the effect of pH on the retention in the range from 4.0 to 9.0 were investigated on both the naked and metal ion chelated columns. Four proteins were quickly separated in 2.0 min with linear gradient elution at a flow rate of 3.0 mL•min -1 by using the synthesized Ni 2＋ -CM-Asp-P GMA/EDMA packings. The separation time was shorter than other immobilized metal affinity chromatography reported in the literature. The Ni 2＋ -CM-Asp-P GMA/EDMA column was further investigated for the rapid separation and purification of copper-zinc superoxide dismutase (Cu,Zn-SOD) from the blood of pig in 3.0 min with only one step. The results obtained were satisfactory.Keywords monodisperse non-porous beads, immobilized metal affinity chromatographic packings, protein, purification www.cjc.wiley-vch.de

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Immobilized iminodiacetic acid (IDA)-type Cu2+-chelating membrane affinity chromatography for purification of bovine liver catalase

Cited by 44 publications

References 25 publications

Material-enhanced laser desorption/ionization (MELDI)—A new protein profiling tool utilizing specific carrier materials for time of flight mass spectrometric analysis

Material-enhanced laser desorption/ionization (MELDI)—A new protein profiling tool utilizing specific carrier materials for time of flight mass spectrometric analysis

Nucleic Acid Separations Utilizing Immobilized Metal Affinity Chromatography

Preparation of Immobilized Metal Affinity Chromatographic Packings by Immobilization of Carboxymethylated Asparate (CM‐Asp) Based on Monodisperse Hydrophilic Non‐porous Beads and Their Application

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