ChromaX®A20 is a protein A affinity chromatography column designed to quickly and accurately determine the antibody concentration (or titer) in cell culture super-natant, which is developed by unique surface treatment technique and coupling technique using recombinant protein A as ligand and polystyrene divinylbenzene (PS-DVB) microspheres with a porous structure as filling carrier.
Characteristics of ChromaX® A20 affinity chromatography column:
Hydrophilic PS-DVB substrate, with the features of good rigidity, low back pres- sure, and fast mass transfer.
Unique hydrophilic treatment technology with low non-specific adsorption en- sures the accuracy of antibody concentration test results.
Ligand specific binding sites, high selectivity ensures the accuracy of concentra- tion determination results.
Stainless steel column for direct use on HPLC.
Fast and efficient, taking only 2 - 5 minutes to complete an analysis.
Good linearity within the wide range of antibody concentration of 0.02 to 10 mg/mL
Adopting alkali-tolerant protein A ligand allows for regenerating the chromato- graphic column with the 0.1 M NaOH solution.
Table 1. Chromatography Column Specifications
Name | ChromaX® A20 |
Column specifications | ID2.1*30 mm; CV=0.1 mL |
Matrix | PS-DVB |
Ligand | Recombinant protein A |
Mean particle size | 20 μm |
Dynamic binding capacity | > 20 mg hIgG / mL chromatography media* |
Recommended flow velocity | 0.5~2.5 mL/min |
Max. Pressure resistance | < 1000 psi |
PH stability range | pH 2 - 10 (common); pH 2 - 13 (CIP) |
CIP cleaning agent | 0.1 M NaOH, etc. |
Operating temperature | 4-40 ℃ |
*Test conditions for dynamic binding capacity: retention for 1 min, when the penetration amount of IgG reaches 10%, IgG loading amount (mg) of unit media volume (mL).
Ordering Information
Product name | Product code | Column tube material | Internal diameter × length | Column volume | Particle size |
ChromaX®A20 | 2821-1111 | SS | 2.1×30 mm | 0.1 mL | 20 μm |
ChromaX® A20 chromatography column shows a long service life by strictly following the instructions for operation. The chromatography column may cause phenomena such as increase in column pressure, decrease in recovery rate and load capacity due to the deposition of samples or miscellaneous proteins, irreversible adsorption of lipids, and other contamination. Regularly cleaning and regenerating the chromatographic column can effectively avoid increasing column pressure and removing residual impurities in the packing, thus achieving the goal of extending the service life of the chromatographic column.
Solvents that can be used for ChromaX® A20 chromatography column include 0.1 M NaOH solution, 1 M acetic acid solution, 1 M acetic acid solution containing 20% ethanol, 20% isopropanol solution, and elution buffer containing 1-2 M NaCl.
Recommended cleaning operation process:
(1) Reverse the chromatographic column into the HPLC system.
(2) Inject the cleaning solvent 2-3 times with the injection volume of 100 μL, (or, rinse the chromatographic column 5-10 CV with the cleaning solvent at the flow velocity of 0.1 mL/min).
(3) Rinse the chromatographic column with the starting buffer at the flow velocity of 0.5-1.0 mL/min.
(4) Insert the chromatographic column forward into the HPLC system.
Reminder: It is recommended to apply reverse rinsing to drive away the particles in the sieve plate of the column head and also to protect the lower half of the column bed from being contaminated. The cleaning solvent needs to be filtered through 0.22 μm membrane before use.
It is recommended to keep it in cold storage at 2 - 8 ℃ after rinsing the chromatography column (20 CV) with the starting buffer. No freezing!
Ensure to tighten the plug screws at both ends of the chromatography column to avoid column drying and cracking!
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