Blue Sepharose 6 Fast Flow

• Cat#No#: Bl-347C
• Product Overview: Blue Sepharose 6 Fast Flow is Cibacron Blue 3G coupled to Sepharose 6 Fast Flow. This resin is well established as adsorbent for albumin and interferon at both laboratory and process scale:
  Designed for large-scale purification of interferon and albumin. Also used for isolating groups of enzymes and removing of albumin. BioProcess resin supported for industrial applications and well-established in approved processes.
  Cibacron Blue 3G covalently coupled to the base matrix allowing for low ligand leakage and high capacity.
• Description: Blue Sepharose 6 Fast Flow is a member of the Cytiva range of affinity chromatography resins for capture and intermediate purification. Blue Sepharose 6 Fast Flow is composed of cross-linked 6% agarose beads modified with Cibacron Blue 3G covalently attached by the triazine coupling method. The blue dye binds many proteins, such as albumin, interferon, lipoproteins and blood coagulation factors. It also binds several enzymes including kinases, dehydrogenases, and most enzymes requiring adenyl-containing cofactors e.g. NAD+.
• Characteristic: High binding capacity.
  Sepharose Fast Flow matrix enables high flow rates.
  Suitable for separation of albumin and interferon.
  Specially developed in cooperation with commercial manufacturers.
• Applications: The most important application areas for Blue Sepharose 6 Fast Flow are the purification of interferon and albumin, as well as albumin removal. The high capacity and high flow rates make the resin suitable for both laboratory and process scale separations
• Maximum operating pressure: 250-400 cm/h, 100 kPa, XK 50/60 column, bed height 25 cm
• Ligand Coupling Method: Triazine
• Packing Column: Blue Sepharose 6 Fast Flow is supplied preswollen in 20% ethanol, 0.1 M KH2PO4, pH 8.0. Decant the solution and replace it with binding buffer. The binding buffer must not contain agents which significantly increase the viscosity, but the column can be equilibrated with viscous buffers at reduced flow rates after packing is completed.
• Matrix: 6% cross-linked agarose
• Particle Size: 45 µm-165 μm
• Average particle size: ~90 µm
• Ligand: Cibacron Blue
• Ligand density: ~ 7 μmol Cibacron Blue/ml medium
• Recommended flow rate: 250-400 cm/h, 100 kPa, XK 50/60 column, bed height 25 cm
• Recommended column height: 25 cm
• Chemical stability: Stable in commonly used aqueous buffers - 70% ethanol, 6 M guanidine hydrochloric, 8 M urea.
• pH working range: 4–12
• CIP stability: 3–13
• Temperature stability: 4°C to 40°C
• Autoclavable: 20 min at 121°C in distilled water
• Storage: 2 to 8°C, 20% Ethanol, 0.1 M Potassium Phosphate buffer, pH 8.0
• Cleaning-in-place: Remove precipitated proteins by washing the column with 4 bed volumes of 0.1 M NaOH solution at a low flow velocity (40 cm/h), followed by washing the column with 3 to 4 bed volumes of 70% ethanol or 2 M potassium thiocyanate.
  Alternatively, wash the column with 2 bed volumes of 6 M guanidine hydrochloride.
  In both cases, wash immediately with at least 5 bed volumes of sterile filtered binding buffer at pH 8.0.
  Remove strongly bound hydrophobic proteins, lipoproteins, and lipids by washing the column with 3 to 4 bed volumes of up to 70% ethanol.
  Alternatively, wash the column with 2 bed volumes of detergent in a basic or acidic solution. Wash at a flow velocity of 40 cm/h. Remove residual detergent by washing with 5 bed volumes of 70% ethanol.
  In both cases, wash immediately with at least 5 bed volumes of sterile filtered binding buffer at pH 8.0.
• Sanitization: 1.Equilibrate the packed column with sterile filtered 70% ethanol.
  2.Allow to stand for 12 h.
  3.Wash with at least 5 bed volumes of sterile filtered buffer.
• Pack size: 50 mL
• BioProcess resin: Yes