High-performance liquid chromatography study of water-soluble ternary polyacrylamide-metal-protein complexes

Dinçer, Betül; Mustafaev, Mamed; Bayülken, Seval

doi:10.1002/(sici)1097-4628(19970705)65:1<37::aid-app5>3.0.co;2-r

Cited by 11 publications

(3 citation statements)

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“…Previously we described the formation of water-soluble and water-insoluble ternary polycomplexes of proteins with synthetic polyelectrolytes (PEs) in the presence of transient metal ions (Fe 3+ , Ba 2+ , Cu 2+ , etc.). [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] Such PE-metalprotein polycomplexes (PMCs) were obtained by simple mixing of solutions of proteins, metal ions, and PEs in water at different pH values. This method consists of the use of small concentrations of metal additives, which promote the PE binding to protein molecule without causing any appreciable changes in the chemical structure of components.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence Study of Cu²⁺-Induced Interaction between Albumin and Anionic Polyelectrolytes

Filenko¹,

Demchenko²,

Mustafaeva³

et al. 2000

Biomacromolecules

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The Cu(2+)-induced complex formation of bovine serum albumin (BSA) with anionic polyelectrolytes (PEs) (polyacrylic acid (PAA), poly(N-isopropylacrylamide) [poly[NIPAAm]], and copolymers of N-isopropylacrylamide (NIPAAm) and acrylic acid) in aqueous solution was studied by a fluorescence technique and high-performance liquid chromatography analysis. The character of the interactions depends on the monomer composition (r = [COOH]/[NIPAAm]), [Cu(2+)]/[PE], and [BSA]/[PE] ratios and solution pH. Two types of ternary polycomplex (polymer + Cu(2+) + BSA) particles are formed depending on the monomer composition r of the copolymer. At r from 1/3 to 1/1, the protein molecules in the structure of ternary polycomplex particles are densely covered by the shell of a polymer coil and practically "fenced off" from the water environment. At r > or = 3/1 ternary polycomplex PAA-Cu(2+)-BSA particles have more friable structures in which protein molecules are practically exposed to the solution. At low polymer concentration, an intrapolymer ternary polycomplex is formed. This complex aggregates to an interpolymer species upon increase in polymer concentration. Fluorescence data indicate that in ternary complex polymer interacts through Cu(2+) ions with BSA preferentially at the site close to the location of "cleft" tryptophan residue. This leads to static quenching of this tryptophan fluorescence. Cu(2+)-induced complex formation is an equilibrium reaction.

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Section: Introductionmentioning

confidence: 99%

Fluorescence Study of Cu²⁺-Induced Interaction between Albumin and Anionic Polyelectrolytes

Filenko¹,

Demchenko²,

Mustafaeva³

et al. 2000

Biomacromolecules

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“…O ponto de carga superficial zero do criogel foi determinado seguindo o procedimento de Davranche et al (2003) e Fioravante (2021). Primeiramente, fragmentos de criogéis de PAAm-CS-GA-AGE-IDA, PAAm-CS-GA-AGE-IDA-Cu 2+ e PAAm-CS-GA-AGE-IDA-Ni 2+ foram secos em estufa.…”

Section: Medição Do Ponto De Carga Superficial Zero Do Criogelunclassified

“…Os criogéis de PAAm-CS-GA, produzidos com a adição de GA (5 ou 10 μL) As cadeias de quitosana são reportadas na literatura por possuírem sítios de coordenação para íons metálicos devido à presença de grupos amino e hidroxila, sendo capazes de adsorver cátions de modo eficiente pelo par de elétrons livres presente no átomo de nitrogênio de seu grupo amino (MONTEIRO; AIROLDI, 1998;WU;TSENG;JUANG, 2001). Além disso, também é relatada a possibilidade de formação de complexos entre as cadeias laterais da poliacrilamida e íons metálicos (SOWWAN et al, 2011;DINÇER;MUSTAFAEV;BAYÜLKEN, 1997) e o agente quelante IDA, alguns autores determinam indiretamente a densidade de IDA imobilizado em matrizes sólidas por meio da quantificação de íons Cu 2+ adsorvidos após a saturação da fase estacionária com solução de sulfato de cobre (ARVIDSSON et al, 2003).…”

Section: Criogéis Sintetizados Com Gaunclassified

Síntese e caracterização de criogéis de poliacrilamida-quitosana com íons metálicos Cu2+ e Ni2+ imobilizados para a purificação de imunoglobulina G humana por IMAC

Sepúlveda Del Rio Hamacek

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Antibodies or immunoglobulins are amongst the proteins that require the most degree of purity for their applications, such as in the diagnostic and treatment of diseases. Current purification methods, in which allow high yields and degree of purity, for those biomolecules can involve affinity chromatography, a technique that is based on the selective interaction between an immobilized ligand and the product of interest. The utilization of those procedures leads to the reduction of the number of additional steps, costs and processing time when compared to other protein purification methods. This work aimed to synthetize and characterize a monolithic stationary phase with adequate properties to be used in immobilized metal ion affinity chromatography to separate immunoglobulin G (hIgG) from human serum. Different conditions of cryopolimerization of the acrylamide, bisacrylamide, allyl-glycidyl-ether in the presence of chitosan chains and the crosslinking agent glutaraldehyde were tested and the cryogel that proved more suitable was selected. The chelating agent iminodiacetic acid (IDA) was covalently immobilized onto the cryogel, which was named as PAAm-CS-GA-AGE-IDA. Then, the adsorption and purification capacities of hIgG by the cryogel in the presence or not of chelated Cu 2+ and Ni 2+ ions were investigated. In the cryogel with chelated Ni 2+ , the sodium phosphate buffer system at pH 7,0 (25 mmol L -1 ) with imidazole (2 mmol L -1 ) proved to be the most promising condition for providing yield of 55% and purity of 88% for hIgG. The breakthrough curve revealed that the cryogel continued to be selective for hIgG under saturation conditions, resulting in 96% of purity. The kinetics experiments evidenced that the equilibrium time of the adsorption is quickly reached. Finally, adsorption isotherms were built and the parameters of the Langmuir and Langmuir-Freundlich models were adjusted by non-linear regression.The Langmuir-Freundlich model represented well the adsorption experimental data, resulting in the maximum capacity equal to 31,19 mg hIgG/g dry cryogel and a dissociation constant in the order of Kd = 10 -7 mol L -1 . The obtained results proved the potential of the cryogel of polyacrylamide and chitosan with immobilized metal ions to separate and purify hIgG from human serum.

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A new approach to human serum albumin chiral stationary phase synthesis and its use in capillary liquid chromatography and capillary electrochromatography

Machtejevas

Maruška

2002

J. Sep. Science

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High-performance liquid chromatography study of water-soluble ternary polyacrylamide-metal-protein complexes

Cited by 11 publications

References 1 publication

Fluorescence Study of Cu²⁺-Induced Interaction between Albumin and Anionic Polyelectrolytes

Fluorescence Study of Cu²⁺-Induced Interaction between Albumin and Anionic Polyelectrolytes

Síntese e caracterização de criogéis de poliacrilamida-quitosana com íons metálicos Cu2+ e Ni2+ imobilizados para a purificação de imunoglobulina G humana por IMAC

A new approach to human serum albumin chiral stationary phase synthesis and its use in capillary liquid chromatography and capillary electrochromatography

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High-performance liquid chromatography study of water-soluble ternary polyacrylamide-metal-protein complexes

Cited by 11 publications

References 1 publication

Fluorescence Study of Cu2+-Induced Interaction between Albumin and Anionic Polyelectrolytes

Fluorescence Study of Cu2+-Induced Interaction between Albumin and Anionic Polyelectrolytes

Síntese e caracterização de criogéis de poliacrilamida-quitosana com íons metálicos Cu2+ e Ni2+ imobilizados para a purificação de imunoglobulina G humana por IMAC

A new approach to human serum albumin chiral stationary phase synthesis and its use in capillary liquid chromatography and capillary electrochromatography

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Fluorescence Study of Cu²⁺-Induced Interaction between Albumin and Anionic Polyelectrolytes

Fluorescence Study of Cu²⁺-Induced Interaction between Albumin and Anionic Polyelectrolytes