Structured Review

GE Healthcare resource rpc
Biophysical characterization of PAS#1(600)-hGH. ( A ) Mass spectrometric analysis of PAS#1(600)-hGH by ESI-MS, confirming the calculated molecular mass of 73121.65 Da as well as precisely monodisperse composition, without any indication of prematurely terminated gene products. ( B ) Hydrophobicity analysis of PASylated hGH by reverse-phase (RP) HPLC. HIC of the original recombinant hGH and its PASylated version was performed on a Resource <t>RPC</t> column using an elution gradient from 2% v/v acetonitrile, 0.065% v/v <t>TFA</t> to 80% v/v acetonitrile, 0.05% v/v TFA. Both profiles show a single homogeneous peak with slightly earlier elution of PAS#1(600)-hGH (51.8% acetonitrile) in comparison with the corresponding unfused protein (52.3% acetonitrile). ( C ) Thermal denaturation of hGH and PAS#1(600)-hGH as determined by far UV CD measurement. Thermal unfolding of a 8.2 µM protein solution in 50 mM K 2 SO 4 , 20 mM K-P i pH 7.5 was followed at a wavelength of 208 nm where maximal spectral change upon unfolding was observed and the normalized unfolded fraction, f(u), was plotted as a function of temperature. The melting temperatures ( T m ) of hGH and PAS#1(600)-hGH were 85.4°C and 88.6°C, respectively.
Resource Rpc, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/resource rpc/product/GE Healthcare
Average 97 stars, based on 16 article reviews
Price from $9.99 to $1999.99
resource rpc - by Bioz Stars, 2020-09
97/100 stars

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1) Product Images from "PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins"

Article Title: PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins

Journal: Protein Engineering, Design and Selection

doi: 10.1093/protein/gzt023

Biophysical characterization of PAS#1(600)-hGH. ( A ) Mass spectrometric analysis of PAS#1(600)-hGH by ESI-MS, confirming the calculated molecular mass of 73121.65 Da as well as precisely monodisperse composition, without any indication of prematurely terminated gene products. ( B ) Hydrophobicity analysis of PASylated hGH by reverse-phase (RP) HPLC. HIC of the original recombinant hGH and its PASylated version was performed on a Resource RPC column using an elution gradient from 2% v/v acetonitrile, 0.065% v/v TFA to 80% v/v acetonitrile, 0.05% v/v TFA. Both profiles show a single homogeneous peak with slightly earlier elution of PAS#1(600)-hGH (51.8% acetonitrile) in comparison with the corresponding unfused protein (52.3% acetonitrile). ( C ) Thermal denaturation of hGH and PAS#1(600)-hGH as determined by far UV CD measurement. Thermal unfolding of a 8.2 µM protein solution in 50 mM K 2 SO 4 , 20 mM K-P i pH 7.5 was followed at a wavelength of 208 nm where maximal spectral change upon unfolding was observed and the normalized unfolded fraction, f(u), was plotted as a function of temperature. The melting temperatures ( T m ) of hGH and PAS#1(600)-hGH were 85.4°C and 88.6°C, respectively.
Figure Legend Snippet: Biophysical characterization of PAS#1(600)-hGH. ( A ) Mass spectrometric analysis of PAS#1(600)-hGH by ESI-MS, confirming the calculated molecular mass of 73121.65 Da as well as precisely monodisperse composition, without any indication of prematurely terminated gene products. ( B ) Hydrophobicity analysis of PASylated hGH by reverse-phase (RP) HPLC. HIC of the original recombinant hGH and its PASylated version was performed on a Resource RPC column using an elution gradient from 2% v/v acetonitrile, 0.065% v/v TFA to 80% v/v acetonitrile, 0.05% v/v TFA. Both profiles show a single homogeneous peak with slightly earlier elution of PAS#1(600)-hGH (51.8% acetonitrile) in comparison with the corresponding unfused protein (52.3% acetonitrile). ( C ) Thermal denaturation of hGH and PAS#1(600)-hGH as determined by far UV CD measurement. Thermal unfolding of a 8.2 µM protein solution in 50 mM K 2 SO 4 , 20 mM K-P i pH 7.5 was followed at a wavelength of 208 nm where maximal spectral change upon unfolding was observed and the normalized unfolded fraction, f(u), was plotted as a function of temperature. The melting temperatures ( T m ) of hGH and PAS#1(600)-hGH were 85.4°C and 88.6°C, respectively.

Techniques Used: Mass Spectrometry, High Performance Liquid Chromatography, Hydrophobic Interaction Chromatography, Recombinant

Related Articles

High Performance Liquid Chromatography:

Article Title: Recombinant Expression, Isotope Labeling and Purification of the Vitamin D Receptor Binding Peptide
Article Snippet: .. Following the cleavage, the entire reaction mixture was loaded onto a Resource RPC column in 1 mL aliquots on HP1100 HPLC system. .. The volume of the cleavage reaction mixture was 10 mL, and the reverse phase chromatography was repeated 10 times.

Article Title: Structural Analysis and Characterization of Lacticin Q, a Novel Bacteriocin Belonging to a New Family of Unmodified Bacteriocins of Gram-Positive Bacteria ▿
Article Snippet: .. This fraction was applied to a 3-ml RESOURCE RPC column (Amersham Biosciences) in an LC-2000Plus high-performance liquid chromatography (HPLC) system (JASCO, Tokyo, Japan). .. An active fraction was eluted in the following manner with a gradient of H2 O-acetonitrile containing 0.1% trifluoroacetic acid at a flow rate of 1 ml/min: 0 to 20 min, 20% to 60% (vol/vol); 20 to 25 min, 60% to 100% (vol/vol); and 25 to 30 min, 100% (vol/vol) acetonitrile.

Article Title: PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins
Article Snippet: .. Reverse-Phase HPLC The purified protein solution in PBS was adjusted to 2% v/v acetonitrile, 0.065% v/v trifluoroacetic acid (TFA) and 1 ml was applied to a Resource RPC 1 ml column (GE Healthcare) equilibrated with buffer A (2% v/v acetonitrile, 0.065% v/v TFA). .. For elution, a 0–100% gradient of buffer B (80% v/v acetonitrile, 0.05% v/v TFA) was applied over 20-bed volumes at a flow rate of 2 ml/min while monitoring protein elution at both 280 and 225 nm (PAS only absorbs in the peptide backbone region due to the absence of aromatic side chains).

Chromatography:

Article Title: Proline Conformation-Dependent Antimicrobial Activity of a Proline-Rich Histone H1 N-Terminal Peptide Fragment Isolated from the Skin Mucus of Atlantic Salmon
Article Snippet: .. Active fractions from the HiPrep SP chromatography were pooled and concentrated by binding to a Resource reversed-phase chromatography (RPC) column (Amersham Biosciences) equilibrated with 0.1% TFA. .. After the bound peptides were washed, they were eluted in 2 to 3 ml of 0.1% TFA in water-2-propanol (1:1) at a flow rate of 1 ml/min.

Purification:

Article Title: Platelet factor 4 regulates megakaryopoiesis through low-density lipoprotein receptor-related protein 1 (LRP1) on megakaryocytes
Article Snippet: .. These proteins were further purified by fast protein liquid chromatography using a Resource RPC FPLC column (Amersham Pharmacia Biotech). .. Purity was assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis followed by silver staining.

Article Title: PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins
Article Snippet: .. Reverse-Phase HPLC The purified protein solution in PBS was adjusted to 2% v/v acetonitrile, 0.065% v/v trifluoroacetic acid (TFA) and 1 ml was applied to a Resource RPC 1 ml column (GE Healthcare) equilibrated with buffer A (2% v/v acetonitrile, 0.065% v/v TFA). .. For elution, a 0–100% gradient of buffer B (80% v/v acetonitrile, 0.05% v/v TFA) was applied over 20-bed volumes at a flow rate of 2 ml/min while monitoring protein elution at both 280 and 225 nm (PAS only absorbs in the peptide backbone region due to the absence of aromatic side chains).

Fast Protein Liquid Chromatography:

Article Title: Platelet factor 4 regulates megakaryopoiesis through low-density lipoprotein receptor-related protein 1 (LRP1) on megakaryocytes
Article Snippet: .. These proteins were further purified by fast protein liquid chromatography using a Resource RPC FPLC column (Amersham Pharmacia Biotech). .. Purity was assessed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis followed by silver staining.

Reversed-phase Chromatography:

Article Title: Screening of DNA Aptamer Against Mouse Prion Protein by Competitive Selection
Article Snippet: .. A three-fold volume of 0.1% (v/v) trifluoroacetic acid (TFA) in distilled water (DW) was added to the oxidized protein solution, and the solution was loaded onto a reverse-phase chromatography column (Resource RPC column, GE Healthcare) pre-equilibrated with 0.1% TFA in DW for solubilization. ..

Article Title: Proline Conformation-Dependent Antimicrobial Activity of a Proline-Rich Histone H1 N-Terminal Peptide Fragment Isolated from the Skin Mucus of Atlantic Salmon
Article Snippet: .. Active fractions from the HiPrep SP chromatography were pooled and concentrated by binding to a Resource reversed-phase chromatography (RPC) column (Amersham Biosciences) equilibrated with 0.1% TFA. .. After the bound peptides were washed, they were eluted in 2 to 3 ml of 0.1% TFA in water-2-propanol (1:1) at a flow rate of 1 ml/min.

Binding Assay:

Article Title: Proline Conformation-Dependent Antimicrobial Activity of a Proline-Rich Histone H1 N-Terminal Peptide Fragment Isolated from the Skin Mucus of Atlantic Salmon
Article Snippet: .. Active fractions from the HiPrep SP chromatography were pooled and concentrated by binding to a Resource reversed-phase chromatography (RPC) column (Amersham Biosciences) equilibrated with 0.1% TFA. .. After the bound peptides were washed, they were eluted in 2 to 3 ml of 0.1% TFA in water-2-propanol (1:1) at a flow rate of 1 ml/min.

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    GE Healthcare resource rpc column
    Elution profile from the Resource <t>RPC</t> column on <t>HPLC.</t> The peak corresponding to VDRBP is marked with an arrow. The acetonitrile gradient is also shown.
    Resource Rpc Column, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 97/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/resource rpc column/product/GE Healthcare
    Average 97 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    resource rpc column - by Bioz Stars, 2020-09
    97/100 stars
      Buy from Supplier

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    Elution profile from the Resource RPC column on HPLC. The peak corresponding to VDRBP is marked with an arrow. The acetonitrile gradient is also shown.

    Journal: Bulletin of the Korean Chemical Society

    Article Title: Recombinant Expression, Isotope Labeling and Purification of the Vitamin D Receptor Binding Peptide

    doi: 10.5012/bkcs.2011.32.12.4337

    Figure Lengend Snippet: Elution profile from the Resource RPC column on HPLC. The peak corresponding to VDRBP is marked with an arrow. The acetonitrile gradient is also shown.

    Article Snippet: Following the cleavage, the entire reaction mixture was loaded onto a Resource RPC column in 1 mL aliquots on HP1100 HPLC system.

    Techniques: High Performance Liquid Chromatography

    Biophysical characterization of PAS#1(600)-hGH. ( A ) Mass spectrometric analysis of PAS#1(600)-hGH by ESI-MS, confirming the calculated molecular mass of 73121.65 Da as well as precisely monodisperse composition, without any indication of prematurely terminated gene products. ( B ) Hydrophobicity analysis of PASylated hGH by reverse-phase (RP) HPLC. HIC of the original recombinant hGH and its PASylated version was performed on a Resource RPC column using an elution gradient from 2% v/v acetonitrile, 0.065% v/v TFA to 80% v/v acetonitrile, 0.05% v/v TFA. Both profiles show a single homogeneous peak with slightly earlier elution of PAS#1(600)-hGH (51.8% acetonitrile) in comparison with the corresponding unfused protein (52.3% acetonitrile). ( C ) Thermal denaturation of hGH and PAS#1(600)-hGH as determined by far UV CD measurement. Thermal unfolding of a 8.2 µM protein solution in 50 mM K 2 SO 4 , 20 mM K-P i pH 7.5 was followed at a wavelength of 208 nm where maximal spectral change upon unfolding was observed and the normalized unfolded fraction, f(u), was plotted as a function of temperature. The melting temperatures ( T m ) of hGH and PAS#1(600)-hGH were 85.4°C and 88.6°C, respectively.

    Journal: Protein Engineering, Design and Selection

    Article Title: PASylation: a biological alternative to PEGylation for extending the plasma half-life of pharmaceutically active proteins

    doi: 10.1093/protein/gzt023

    Figure Lengend Snippet: Biophysical characterization of PAS#1(600)-hGH. ( A ) Mass spectrometric analysis of PAS#1(600)-hGH by ESI-MS, confirming the calculated molecular mass of 73121.65 Da as well as precisely monodisperse composition, without any indication of prematurely terminated gene products. ( B ) Hydrophobicity analysis of PASylated hGH by reverse-phase (RP) HPLC. HIC of the original recombinant hGH and its PASylated version was performed on a Resource RPC column using an elution gradient from 2% v/v acetonitrile, 0.065% v/v TFA to 80% v/v acetonitrile, 0.05% v/v TFA. Both profiles show a single homogeneous peak with slightly earlier elution of PAS#1(600)-hGH (51.8% acetonitrile) in comparison with the corresponding unfused protein (52.3% acetonitrile). ( C ) Thermal denaturation of hGH and PAS#1(600)-hGH as determined by far UV CD measurement. Thermal unfolding of a 8.2 µM protein solution in 50 mM K 2 SO 4 , 20 mM K-P i pH 7.5 was followed at a wavelength of 208 nm where maximal spectral change upon unfolding was observed and the normalized unfolded fraction, f(u), was plotted as a function of temperature. The melting temperatures ( T m ) of hGH and PAS#1(600)-hGH were 85.4°C and 88.6°C, respectively.

    Article Snippet: Reverse-Phase HPLC The purified protein solution in PBS was adjusted to 2% v/v acetonitrile, 0.065% v/v trifluoroacetic acid (TFA) and 1 ml was applied to a Resource RPC 1 ml column (GE Healthcare) equilibrated with buffer A (2% v/v acetonitrile, 0.065% v/v TFA).

    Techniques: Mass Spectrometry, High Performance Liquid Chromatography, Hydrophobic Interaction Chromatography, Recombinant