TSKgel Protein A-5PW

The TSKgel Protein A-5PW column has been designed for the rapid separation and robust quantification of a variety of antibodies. Monoclonal antibodies from harvested cell culture media can be captured and accurately quantitated in less than 2 minutes per injection. The column can be used for more than 2,000 injections without regeneration or cleaning.

Packed with hydroxylated methacrylic polymer beads, the TSKgel Protein A-5PW column is designed with a high degree of crosslinking, which allows high flow rate for chromatography while still maintaining chromatographic efficiency, peak width and resolution. The recombinant protein A ligand is a code-modified hexamer of the C domain. An enhanced rProtein A ligand is bound to the TSKgel 5PW base bead via multipoint attachment resulting in excellent base stability in 0.1 mol/L NaOH.

The wide range loading capacity of the TSKgel Protein A-5PW column can accurately determine the titer of mAb at various stages of cell culture media processing. The low level of protein A leaching makes this column a good candidate for small scale purification of mAbs for initial characterization.

P/N	Description	Particle Size (µm)	Housing Material	ID (mm)	Length (cm)	Price ($)
0023483	TSKgel Protein A-5PW	20	PEEK	4.6	3.5	1,287	Inquire

Product Attributes

Pore size (mean):	100 nm
Particle size:	20 µm
pH stability:	2.0 - 12.0
Exclusion limit:	1,000 kDa
Ligand:	Recombinant protein A, hexamer of C domain

Affinity for Various Antibodies

Species	Subclass	Protein A ligand of Protein A-5PW	Native Protein A
Human	IgG₁ IgG₂ IgG₃ IgG₄	+++++ +++++ - +++++	++++ ++++ - ++++
Mouse	IgG₁ IgG_2a IgG_2b IgG₃	++++ +++++ +++++ ++++	+ ++++ +++ ++
Rat	IgG₁ IgG_2a IgG2b IgG2c	++++ - +++ ++++	- - - -
Goat	IgG_S	++++	-
Chicken	IgY	-	-
Rabbit	IgG	+++++	++++

Rapid Separation of IgG from Impurities

Column:	TSKgel Protein A-5PW, 20 µm, 4.6 mm ID × 3.5 cm
Binding buffer:	20 mmol/L sodium phosphate buffer, pH 7.4
Elution buffer:	20 mmol/L sodium phosphate buffer, pH 2.5
Stepwise gradient:	0 - 0.5 min: binding buffer 0.5 - 1.1 min: elution buffer 1.1 - 2.0 min: binding buffer
Flow Rate:	2 mL/min
Detection:	UV @ 280 nm
Sample:	20 µL of CHO cell culture supernatant spiked with polyclonal IgG (0.5 mg/mL)

For additional applications, please see our applications database.

Durability and Dynamic Range

The TSKgel Protein A-5PW column was subjected to a linearity analysis test. Purified IgG was initially injected onto the column with subsequent injections of IgG made at different volumes. The column was then used up to 2,009 injections without being cleaned. A linearity analysis test was then repeated. No significant change in the calibration curve for IgG was seen. The column still maintained its high loading capacity with an excellent linearity (R²=0.9999).

Wide Range of Loading Concentrations of Purified IgG

Column:	TSKgel Protein A-5PW, 20 µm, 4.6 mm ID × 3.5 cm (PEEK)
Binding and washing buffer:	20 mmol/L sodium phosphate buffer, pH 7.4
Elution buffer:	20 mmol/L sodium phosphate buffer, pH 2.5 Note: IgG can also be eluted with 12 mmol/L HCL, 20-100 mmol/L citric acid, pH 2.5-3.5, 20-100 mmol/L glycine, pH 2.5-3.5, 5-10% acetic acid
Stepwise gradient:	0 - 0.5 min: binding buffer 0.5 - 1.1 min: elution buffer 1.1 - 2.0 min: binding buffer
Flow rate:	2 mL/min
Detection:	UV @ 280 nm
Sample:	IgG

Leaching Analysis

Resin	Concentration (mg/mL)	Protein A (ppm)
TSKgel Protein A-5PW	0.538	2.3
Competitor Protein A	0.508	10.8

An analysis of an IgG sample was run on both a TSKgel Protein A-5PW, 20 μm, 4.6 mm ID × 3.5 cm column and a similar competitive 20 μm protein A, 4.6 mm ID × 5 cm column. The table shows that the protein A ligand from the TSKgel Protein A-5PW column has 2.3 ppm of Protein A leaching, which is many folds lower than the competitor’s protein A column. This data suggests that the coupling process of protein A onto the TSKgel 5PW base bead is better than its competitor’s coupling process.

Varying Flow Rates to Capture IgG

Column:	TSKgel Protein A-5PW, 20µm, 4.6 mm ID × 3.5 cm (PEEK)
Binding and washing buffer:	20 mmol/L sodium phosphate buffer, pH 7.4
Elution buffer:	20 mmol/L sodium phosphate buffer, pH 2.5 Note: IgG can also be eluted with 12 mmol/L HCL, 20-100 mmol/L citric acid, pH 2.5-3.5, 20-100 mmol/L glycine, pH 2.5-3.5, 5-10% acetic acid
Stepwise gradient:	0 - 0.5 min: binding buffer 0.5 - 1.1 min: elution buffer 1.1 - 2.0 min: binding buffer
Flow rate:	2 mL/min
Detection:	UV @ 280 nm
Sample:	IgG

Code #	Description	Literature Type
AN096	Fast monoclonal antibody titer determination with TSKgel Protein A-5PW column	Application Note
BR09	Solutions for Monoclonal Antibody Analysis and Characterization	Brochure
DS1242	TSKgel Protein A-5PW Products	Operating Conditions and Specifications
IM01	TSKgel Column Instruction Manual	Instruction Manual
LCAT08	TSKgel U/HPLC Columns 2019 Product Guide -- Protein A Columns	Product Catalog
PO44	TSKgel Protein A-5PW Column	Product Overview
TP218	High Throughput and Robust Method of Analysis of a Variety of Immunoglobulin G	Technical Presentation
TP220	High Throughput and Robust Method of Analysis of a Variety of Immunoglobulin G (IgGs) Using an Analytical Recombinant Protein A Affinity Chromatography Column	Technical Presentation