Purification of human adenosine deaminase for the preparation of a reference material

Human adenosine deaminase as an allosteric modulator of human A1adenosine receptor: abolishment of negative cooperativity for [3H](R)‐pia binding to the caudate nucleus

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside‐Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al. (2000) for ADA from human erythrocytes.…”

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside-Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al (2000) for ADA from human erythrocytes. The enzyme preparation obtained by this short procedure exhibited a specific activity of 442 U/mg protein at 37°C, analogous to the naturally occurring ADA from human T lymphoblasts (Richard et al 2002).…”

Human adenosine deaminase as an allosteric modulator of human A1adenosine receptor: abolishment of negative cooperativity for [3H](R)‐pia binding to the caudate nucleus

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside‐Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al. (2000) for ADA from human erythrocytes.…”

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside-Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al (2000) for ADA from human erythrocytes. The enzyme preparation obtained by this short procedure exhibited a specific activity of 442 U/mg protein at 37°C, analogous to the naturally occurring ADA from human T lymphoblasts (Richard et al 2002).…”

Molecular dynamics simulation studies of the conformation and lateral mobility of a charged adsorbate biomolecule: Implications for estimating the critical value of the radius of a pore in porous media

“…The large-scale production of diagnostics and biotherapeutics has resulted from the important advances that have occurred in the past twenty years in the fields of molecular biology and separation science [1][2][3][4][5][6][7][8][9][10][11]. In downstream purification of bioactive macromolecules, the capture step serves [1][2][3][4][5][6] the dual purpose of removing the bulk of impurities and in concentrating the product of interest, and, because of this, ion-exchange chromatography (IEC) has become a very important separation method in downstream purification for the isolation [1][2][3][4][5][6][7][8][9][10][11] of peptides, enzymes, and proteins.…”

“…In downstream purification of bioactive macromolecules, the capture step serves [1][2][3][4][5][6] the dual purpose of removing the bulk of impurities and in concentrating the product of interest, and, because of this, ion-exchange chromatography (IEC) has become a very important separation method in downstream purification for the isolation [1][2][3][4][5][6][7][8][9][10][11] of peptides, enzymes, and proteins. Furthermore, the mass transport and interaction (i.e., adsorption/desorption) mechanisms involved in the isolation of biologically active charged macromolecules have also to be considered, because of their important role in systems involving food processing, bacterial cell adhesion, and in the development of biomaterials and biosensors [11][12][13][14][15][16].…”

Human adenosine deaminase as an allosteric modulator of human A1adenosine receptor: abolishment of negative cooperativity for [3H](R)‐pia binding to the caudate nucleus

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside‐Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al. (2000) for ADA from human erythrocytes.…”

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside-Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al (2000) for ADA from human erythrocytes. The enzyme preparation obtained by this short procedure exhibited a specific activity of 442 U/mg protein at 37°C, analogous to the naturally occurring ADA from human T lymphoblasts (Richard et al 2002).…”

Molecular dynamics simulation studies of the conformation and lateral mobility of a charged adsorbate biomolecule: Implications for estimating the critical value of the radius of a pore in porous media

“…The large-scale production of diagnostics and biotherapeutics has resulted from the important advances that have occurred in the past twenty years in the fields of molecular biology and separation science [1][2][3][4][5][6][7][8][9][10][11]. In downstream purification of bioactive macromolecules, the capture step serves [1][2][3][4][5][6] the dual purpose of removing the bulk of impurities and in concentrating the product of interest, and, because of this, ion-exchange chromatography (IEC) has become a very important separation method in downstream purification for the isolation [1][2][3][4][5][6][7][8][9][10][11] of peptides, enzymes, and proteins.…”

“…In downstream purification of bioactive macromolecules, the capture step serves [1][2][3][4][5][6] the dual purpose of removing the bulk of impurities and in concentrating the product of interest, and, because of this, ion-exchange chromatography (IEC) has become a very important separation method in downstream purification for the isolation [1][2][3][4][5][6][7][8][9][10][11] of peptides, enzymes, and proteins. Furthermore, the mass transport and interaction (i.e., adsorption/desorption) mechanisms involved in the isolation of biologically active charged macromolecules have also to be considered, because of their important role in systems involving food processing, bacterial cell adhesion, and in the development of biomaterials and biosensors [11][12][13][14][15][16].…”

Human adenosine deaminase as an allosteric modulator of human A1adenosine receptor: abolishment of negative cooperativity for [3H](R)‐pia binding to the caudate nucleus

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside‐Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al. (2000) for ADA from human erythrocytes.…”

“…By optimizing the elution conditions (a temperature of 30°C and a high substrate concentration in the equilibration buffer), retained ADA can be efficiently recovered from the purine riboside-Sepharose column. The final enzyme concentration as well as the yield (see Table 1) is much higher than those obtained by Bota et al (2000) for ADA from human erythrocytes. The enzyme preparation obtained by this short procedure exhibited a specific activity of 442 U/mg protein at 37°C, analogous to the naturally occurring ADA from human T lymphoblasts (Richard et al 2002).…”

Molecular dynamics simulation studies of the conformation and lateral mobility of a charged adsorbate biomolecule: Implications for estimating the critical value of the radius of a pore in porous media

“…The large-scale production of diagnostics and biotherapeutics has resulted from the important advances that have occurred in the past twenty years in the fields of molecular biology and separation science [1][2][3][4][5][6][7][8][9][10][11]. In downstream purification of bioactive macromolecules, the capture step serves [1][2][3][4][5][6] the dual purpose of removing the bulk of impurities and in concentrating the product of interest, and, because of this, ion-exchange chromatography (IEC) has become a very important separation method in downstream purification for the isolation [1][2][3][4][5][6][7][8][9][10][11] of peptides, enzymes, and proteins.…”

“…In downstream purification of bioactive macromolecules, the capture step serves [1][2][3][4][5][6] the dual purpose of removing the bulk of impurities and in concentrating the product of interest, and, because of this, ion-exchange chromatography (IEC) has become a very important separation method in downstream purification for the isolation [1][2][3][4][5][6][7][8][9][10][11] of peptides, enzymes, and proteins. Furthermore, the mass transport and interaction (i.e., adsorption/desorption) mechanisms involved in the isolation of biologically active charged macromolecules have also to be considered, because of their important role in systems involving food processing, bacterial cell adhesion, and in the development of biomaterials and biosensors [11][12][13][14][15][16].…”