Semba Biosciences’ Octave™ Chromatography System is suitable for use with biological samples including crude cell lysates, tissue culture fluids, serum samples.

The Octave System is compatible with a wide range of columns and biochromatography media. Column sizes vary depending on the required separation scale. For example, 1-ml and 5-ml disposable columns are ideal for use with the Octave System for purifying 3 to 15 grams per day, respectively. In general, columns packed with any polymeric resin (e.g. cross-linked agarose, methacrylate, polyacrylamide) having a particle size greater than 35 microns are compatible with the Octave Systems. In all cases, as with any high performance chromatography system, samples should be free of particulate matter. Filtration of samples through a 0.45 micron membrane is usually sufficient.

Octave™ Ni-NTA SF Columns

The Octave Ni-NTA SF Columns are designed for purification of histidine-tagged recombinant proteins or other metal-binding proteins by immobilized metal affinity chromatography (IMAC).

Columns in 1-ml or 5-ml sizes are packed with Ni-NTA Superflow, the most-cited resin for IMAC purification of histidine-tagged proteins from any expression system under native or denaturing conditions. The Superflow agarose resin is covalently coupled to nitriloacetic acid (NTA) and pre-charged with nickel. NTA, which has four chelation sites, binds nickel and other metals more tightly than systems that only have three sites available for interaction with metal ions. The extra chelation site prevents metal leaching and results in higher purity protein preparations, making this resin ideal for continuous SMBC processes involving multiple cycles of purification without intermediate stripping and recharging. Superior mechanical stability and outstanding flow characteristics of the Superflow matrix also enable efficient continuous purification of target proteins in Step SMBC Mode.

Octave Ni-NTA SF columns are compatible with a wide range of buffer components, including strong denaturants, detergents, and reducing agents (up to 20 mM 2-mercaptoethanol). The columns can be quickly and easily connected to the Octave Chromatography System through 10-32 F to M6 F adapters (available separately).

Ni-NTA Step mode enolase gel labeled

Step SMBC IMAC purification of histidine-tagged protein with Octave™ Ni-NTA SF Columns

Histidine-tagged human enolase was expressed in E. coli. A bacterial lysate was prepared by standard methods and applied to 1-ml Octave Ni-NTA SF Columns on the Octave 10 System in a 2-2-2-2 Step SMBC configuration. Columns were equilibrated in Bind Buffer (50 mM Na phosphate, pH 8.0, 300 mM NaCl), washed in Bind Buffer plus 20 mM imidazole (SembaChrom grade), and eluted with Bind Buffer plus 250 mM imidazole. M, Markers (10-225 kDa); Feed, E. coli lysate containing recombinant His-enolase; SMB, SMBC-purified product. The purified target protein can be seen as a single band at 55 kDa.

Octave™ Ni-NTA SF Column Specs

Binding capacity: Up to 50 mg/ml (up to 2500 nmol @ ~20 kDa)
Support: Superflow
Bead structure: Highly cross-linked, 6% agarose
Bead size: 60–160 µm
Column dimensions, 1 ml: 6.7 mm I.D. x 28 mm L
Column dimensions, 5 ml: 14.7 mm I.D. x 29.8 mm L
Column housing material Polypropylene

Octave™ MabCapture® A Columns

The Octave MabCapture A Columns are designed for rapid Protein A affinity purification of antibodies using advanced Perfusion Chromatography® technology.

Columns in 1-ml and 5-ml sizes are packed with POROS® MabCapture A media. The patented POROS polymer support enables use of much higher flow rates than conventional media while retaining high binding capacity, resulting in faster separations. The rigid particles are also less susceptible to compression and back pressure limitations even when operating at flow rates from 1000 to 4500 cm/h, permitting efficient operation over a broad range of conditions.

Proprietary surface chemistry facilitates efficient Protein A coupling with low leaching through many cycles of binding and elution, and is compatible with standard sodium hydroxide sanitation. The unique macroporous matrix allows flow rates up to 20 ml/min (1-ml columns) and 80 ml/min (5-ml columns), speeding the purification procedure and increasing throughput. The columns are quickly and easily connected to the Octave Chromatography System through 10-32 F to M6 F adapters (available separately). The columns can also be used for automated purification on single-column liquid chromatography systems and for manual purification using a syringe.

poros

Continuous multicolumn Protein A affinity chromatography of monoclonal antibody with Octave™ MabCapture™ A Columns

Monoclonal antibody was produced in mammalian cell culture. The culture fluid was concentrated, diafiltered, and applied to 8 x 1-ml Octave MabCapture A Columns on the Octave 10 System in a 2-2-2-2 Step SMBC configuration. The indicated fractions were analyzed by SDS-polyacrylamide gel electrophoresis and Coomassie blue staining. Feed, concentrated tissue culture fluid; Raff., flow-through (raffinate); SMB Purified, duplicate loads of SMBC product; M: Markers 10-225 kDa. mAb heavy chain (HC) and light chain (LC) bands are indicated.

Octave™ MabCapture A Column Specs

Binding capacity: >45 mg/ml
Support: Cross-linked Poly (styrene-divinylbenzene)
Ligand: Recombinant Protein A
Bead size: 45 micron
Column dimensions, 1 ml: 6.7 mm I.D. x 28 mm L
Column dimensions, 5 ml: 14.7 mm I.D. x 29.8 mm L
Column housing material Polypropylene

Octave™ MabCapture A Product Literature

Made with POROS® MabCapture® A media from Life Technologies Corporation. POROS®and MabCapture® are Registered Trademarks of Life Technologies Corporation