Bio-Scale™ MT Columns Instruction Manual Catalog numbers 751-0081 751-0083 751-0085 751-0087
Table of Contents Section 1 Characteristics of Bio-Scale MT2, MT5, MT10, and MT20 Columns ......................... 1 1.1 Introduction ........................................................... 1 1.2 Column Description ............................................... 1 1.3 Column Specifications ........................................... 2 Section 2 Use of the Bio-Scale MT Columns ..................... 2 2.1 Column Disassembly ............................................. 4 2.
Section 1 Characteristics of the Bio-Scale MT2, MT5, MT10, and MT20 Columns 1.1 Introduction The Bio-Scale MT columns have been designed specifically to provide high resolution chromatography of bio-molecules when packed with any suitable chromatography media. Typical packings might include ion exchange, hydrophobic interaction, hydroxyapatite, or affinity chromatography media.
1.3 Column Specifications All wetted parts are bio-compatible (Tefzel, polypropylene, polyphenyl sulfide, borosilicate glass and buna-N rubber) and resistant to cleaning and sanitation solvents including 1.0 M NaOH and 25% acetic acid. The column can be used with most common aqueous buffers and certain organic solvents including alcohols and acetonitrile. Do not use halogenated hydrocarbons, aromatic solvents, or tetrahydrofuran (THF). A detailed solvent compatibility table is given in Section 4. Table 1.
Section 2 Use of Bio-Scale MT Columns Upper end fitting (1) Lock nut (2) Upper end cap (3) Protective shield (4) Upper bed support (5) Distribution screen (6) Frit (7) FLOW Glass column (8) Lower bed support (9) Lower end cap (10) Lower end fitting (11) Fig. 1. Diagram of Bio-Scale MT Column.
2.1 Column Disassembly (part given are in parenthesis) 1. Unscrew and remove the upper (1) and lower (11) end fittings. 2. Unscrew the bottom end cap (10) from the protective shield (4). 3. Withdraw the borosilicate glass column assembly (8) from the protective shield. 4. Remove the upper (5) and lower (9) bed supports. 5. If the column is packed, remove the upper and lower distribution screens (7) and frits (6) using the frit removal tool. Do not reuse these parts.
. Gently turn the lock nut until the upper bed support is in the desired position. 10. Attach upper (1) and lower (11) end fittings. 2.3 Column Packing Always follow the manufacturer’s instructions for packing a particular type of media. Generally, slurry packing will give better chromatographic performance than dry packing. The following guidelines may be useful: Dry packing Particle size Ž 20 µm Slurry packing Particle size < 20 µm Dry Packing Assemble the column according to the following figure.
5. Attach the column to a pump and start flow at 50% of the desired operating flow rate. 6. When all the air is expelled from the column, increase the flow rate to 20% higher than the desired operating flow rate for at least 5 column volumes. 7. Stop the flow and lower the upper bed support via the lock nut to remove any head space that might have formed between the frit and the top of the bed. 8. Equilibrate the column at the desired flow rate prior to sample application.
8. Pass 5 bed volumes of buffer through the column at a flow rate 20% higher than the operating flow rate. 9. If necessary, stop the flow and lower the upper bed support via the lock nut to remove any head space that might have formed between the frit and the top of the bed. 10. Equilibrate the column at the desired flow rate. 2.4 Connections to Chromatography Systems Bio-Scale MT columns can be used with any chromatography system that can provide the appropriate flow rate and pressure.
Section 3 Care of the Bio-Scale MT Columns 3.1 Frit Removal The top frit may need to be replaced after extensive column use or if increasing backpressures are noticed. Each column is supplied with a frit removal tool, polyethylene frit, and distribution screen. The screen assists in distributing the sample over the entire column surface and also acts as a pre-filter. The screen should be replaced every time the frit is changed, refer to Figure 1. 1. Remove the upper and lower end fittings from the column 2.
Section 4 Solvent Compatibility Table Table 2. Solvent Compatibility. Solvents Acetic acid (50%) Acetonitrile Butanol Citric acid Dimethyl formamide (DMF) Dimethylsulfoxide (DMSO) Ethanol Elthylenediamine tetraacetic acid (EDTA) Formaldehyde (40%) Formic acid Glycerol Goods buffers Guanadine-HCl (6 M) Hydrocloric acid (2 M) Isopropanol Lactic acid (85%) Methanol SDS Sodium acetate (sat.) Sodium hydroxide (1 M) Sodium thiocyanate (3 M) Sulfuric acid (dilute) Trifluoroacetic acid (0.
Section 5 Product Information Catalog Number Product Description Bed Dimension, mm Bed Volume, ml Pressure Limit, psi 751-0081 Bio-Scale MT2 7 x 52 2 1.000 751-0083 Bio-Scale MT5 10 x 64 5 750 751-0085 Bio-Scale MT10 12 x 88 10 600 751-0087 Bio-Scale MT20 15 x 113 20 500 All columns come with 5 extra frits, 5 distribution screens, frit removal tool, 2 o-rings, 2 1/4 x 28 fittings, 2 M6 fittings, and 2 10-32 fittings.
Bio-Scale MT Column Instruction Manual 11
Bio-Rad Laboratories, Inc. Web site www.bio-rad.
