TSKgel Protein A-5PW chromatographic column

TSKgel Protein A-5PW chromatographic column

The TSKgel Protein A-5PW chromatographic column is designed specifically for rapid separation and accurate quantification of various antibodies. Capture and accurately determine the amount of monoclonal antibodies from the harvested cell culture medium, and each injection analysis can be completed in less than 2 minutes. This chromatographic column can perform over 2000 injection analyses without the need for regeneration or cleaning.

The TSKgel Protein A-5PW chromatographic column is filled with highly cross-linked hydroxylated methyl methacrylate polymer beads, allowing for the use of high flow rates in chromatographic analysis while maintaining chromatographic efficiency, peak width, and separation. The recombinant protein A ligand is a codon modified C-domain hexamer. This enhanced recombinant protein A ligand is bonded to TSKgel 5PW matrix microspheres through multi-point attachment, resulting in excellent stability in 0.1 mol/L NaOH solution.

The TSKgel Protein A-5PW chromatographic column has a wide sample loading range and can accurately determine the potency of monoclonal antibodies in different stages of cell culture medium treatment. Its low protein A dropout rate makes it an ideal choice for small-scale monoclonal antibody purification for preliminary characterization.

TSKgel Protein A-5PW is an affinity (AFC) chromatography column that introduces recombinant Protein A functional groups onto the surface of TSKgel G5000PW filler in a porous hydrophilic polymer matrix and fills it into PEEK column tubes.

Due to its specific binding to the Fc region of IgG, Protein A can selectively adsorb and separate IgG from samples such as cell culture supernatants containing IgG and impurities. In addition, calibration was performed using IgG of known concentration

The curve can quantify IgG in samples with unknown concentrations.

The matrix, functional groups, and functional group bonding method of TSKgel Protein A-5PW chromatographic column are the same as those of bulk packing TOYOPEARL AF rProtein A HC-650F. Therefore, this chromatographic column has similar separation selectivity performance to TOYOPEARL AF rProtein A HC-650F.

Due to the large particle size and high mechanical strength of the matrix, this chromatographic column can be used under high flow rate conditions and can also be used for high-throughput analysis of IgG. Moreover, due to the relatively small swelling effect of the filler caused by solvent changes, it can be added in the mobile phase

Add organic solvents.

In addition, the chromatographic column also uses functional groups with high chemical stability and alkali resistance, so NaOH aqueous solution can be used for cleaning. When cleaning the chromatography column with NaOH aqueous solution, please inject 0.1 mol/L NaOH aqueous solution 0.5 through the injection valve

〜2 mL。

2-2. Standard separation conditions

Most IgG is adsorbed by Protein A under neutral conditions (pH 7.0-7.5) and eluted under acidic conditions (pH 2.5-3.5). Therefore, when using TSKgel Protein A-5PW to separate antibodies, two different pH solutions are usually used

Ladder gradient elution method.

The mobile phase for adsorption and elution generally uses phosphate buffer, but low pH citrate buffer can also be used as the mobile phase. After pre equilibrating the chromatographic column with adsorption buffer (equilibrating with buffer at least 5 times the column volume), inject the sample

Afterwards, only IgG was adsorbed onto the column. Afterwards, switch to the elution mobile phase to elute IgG.

High concentration salts (such as 0.15 mol/L NaCl, etc.) can be added to the mobile phase, but IgG with high hydrophobicity is prone to non-specific adsorption on the chromatographic column, and more attention should be paid to this. In addition, when analyzing IgG with high hydrophobicity

Adding water-soluble organic solvents such as ethanol or arginine (0.2mol/L) to the equilibrium solution and eluent can effectively inhibit non-specific adsorption. However, the above situation may cause baseline fluctuations due to the replacement of the mobile phase, so before injection, please make sure to

Change the mobile phase and confirm.

Example of mobile phase

Example 1

Mobile phase A: 20 mmol/L sodium phosphate buffer, pH 7.4

Mobile phase B: 20 mmol/L sodium phosphate buffer, pH 2.5

Example 2

Mobile phase A: 20 mmol/L sodium phosphate buffer, pH 7.4/ethanol=90/10 (v/v)

Mobile phase B: 20 mmol/L sodium phosphate buffer, pH 2.5/ethanol=90/10 (v/v)

Example 3

Mobile phase A: 20 mmol/L sodium phosphate buffer, pH 7.4

Mobile phase B: 50 mmol/L sodium citrate buffer, pH 2.5

The high-performance affinity chromatography column TSKgel Protein A-5PW has good durability and can be cleaned with alkaline solution. It is suitable for rapid and high-precision quantification of IgG in cell culture media containing high impurities. In addition, IgG has a wide quantitative range and can be used to analyze low concentration samples to high concentration samples in the early stages of culture.

TSKgel Protein A-5PW Product Specifications and Ordering Information

Specifications

projectparameter

aperture100 nm

particle size20 µm

ph stability2.5 - 12

Exclusion limit1,000 kDa

ligandRecombinant protein A (C domain hexamer)

Ligand density0.9-1.0 g/L gel

Order Information

Product Number (P/N)descriptionsize

0023483TSKgel Protein A-5PW chromatographic column4.6 mm inner diameter x 3.5 cm length

TSKgel Protein A-5PW chromatographic column

Nantong Hairuo Chemical Co., Ltd