To estimate product purity, size exclusion chromatography was performed. For HPLC analytics, we used a Dionex UltiMate 3000 HPLC system equipped with a diode array detector (Thermo Fisher Scientific). The running buffer was a 50 mM potassium phosphate buffer with 150 mM NaCl, pH 7.0 (Merck KGaA). The buffer was filtered through 0.22‐μm filters (Merck KGaA) and degassed. We applied 20 μl of the filtered sample to a TSKgel® G3000SWXL HPLC column (5 μm, 7.8 × 300 mm2) in combination with a TSKgel SWXL Guard column (7 μm, 6.0 × 40 mm2; Tosoh). The absorbance at 280 nm was recorded, and the results were evaluated with the Chromeleon™ 7 software (Thermo Fisher Scientific). The antibody purity was calculated as the ratio of the monomer peak area (retention time: 21.2 min) to the sum of all peak areas, based on the 280 nm signal.
G3000swxl hplc column
Manufactured by Tosoh
Sourced in Japan, United States
The G3000SWXL HPLC Column is a size exclusion chromatography column designed for the analysis of macromolecules such as proteins, peptides, and polymers. It features a silica-based packing material with a pore size of 300 Å, which allows for the separation of molecules within a molecular weight range of 1,000 to 300,000 Daltons. The column dimensions are 7.8 mm x 300 mm.
Automatically generated - may contain errors
Lab products found in correlation
Chromeleon 7, by Thermo Fisher Scientific (3 mentions)
Tskgel swxl guard column, by Tosoh (3 mentions)
Dionex ultimate 3000 hplc system, by Thermo Fisher Scientific (2 mentions)
0.22 μm filter, by Merck Group (1 mentions)
0.22 m filter, by Merck Group (1 mentions)
Gswp04700, by Merck Group (1 mentions)
L 1 nacl, by Merck Group (1 mentions)
Dionex ultimate 3000, by Thermo Fisher Scientific (1 mentions)
3 protocols using g3000swxl hplc column
1
Antibody Purity Estimation by SEC-HPLC
2
Antibody Purity Analysis via HPLC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We used size exclusion chromatography to estimate product purity. For HPLC analytics, we used a Dionex UltiMate 3000 HPLC system equipped with a diode array detector (Thermo Fisher Scientific). The running buffer was a 50 mM potassium phosphate buffer with 150 mM NaCl, pH 7.0 (Merck KGaA). The buffer was filtered through 0.22 µm filters (Merck KGaA) and degassed. We applied 20 µL of the filtered sample to a TSKgel® G3000SWXL HPLC Column (5 µm, 7.8 × 300 mm) in combination with a TSKgel SWXL Guard Column (7 µm, 6.0 × 40 mm; Tosoh, Tokyo, Japan). The absorbance at 280 nm was recorded, and the results were evaluated with the Chromeleon™ 7 software (Thermo Fisher Scientific). The antibody purity was calculated as the ratio of the monomer peak area (retention time 21.2 min) to the sum of all peak areas, based on the 280 nm signal.
3
Antibody Characterization by Size Exclusion Chromatography
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Size exclusion chromatography was used to assess antibody yield, purity, and the amount of HMWI. High‐performance liquid chromatography was carried out by isocratic elution on a Dionex UltiMate 3000 HPLC system equipped with a diode array detector (Thermo Scientific, Waltham, MA, USA). The running buffer was 50 mmol L‐1 sodium phosphate buffer with 150 mmol L‐1 NaCl (Sigma‐Aldrich) at a pH of 7.0, prepared with 0.22 µm filtration (GSWP04700, Merck KGaA). 10 µL of a 0.2 µm vacuum‐filtered sample (0.2 µm GHP AcroPrep™ 96 filter plate; Pall Life Sciences, Ann Arbor, MI, USA) was applied to a TSKgel® G3000SWXL HPLC Column (5 µm, 7.8 × 300 mm) with a TSKgel SWXL Guard Column (7 µm, 6.0 × 40 mm; Tosoh, Tokyo, Japan). Chromeleon™ 7 software (Thermo Scientific) was used to monitor the signals at 215 nm (for HMWI) and 280 nm (for purity and yield). Product purity was defined by the ratio of product peak area (monomer) to sum of all peak areas. Product yield was calculated dividing the product peak area (monomer) of the flocculated material by the product peak area (monomer) of the cell culture broth.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!