SEC was performed as described,30 using a Shimadzu Prominence ultra-fast liquid chromatography HPLC system. 10 µL of mAb (10 µg total protein) was injected and separated by a TSKgel G4000SWXL column (8 µm particle size, 7.8 mm ID × 30 cm) with the corresponding guard column operated at ambient temperature (Tosoh Biosciences) using a 30-minute run time. Gel filtration molecular weight standards (Bio-Rad, Hercules, CA) were injected as controls. Data were analyzed using LC-Solutions software (Shimadzu, Kyoto, Japan).
Prominence ultra fast liquid chromatography hplc system
Manufactured by Shimadzu
The Prominence ultra-fast liquid chromatography HPLC system is a high-performance liquid chromatography instrument manufactured by Shimadzu. It is designed for the separation, identification, and quantification of complex mixtures of chemical compounds.
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Lab products found in correlation
Corresponding guard column, by Tosoh (2 mentions)
Lc solutions software, by Shimadzu (1 mentions)
Gel filtration molecular weight standard, by Bio-Rad (1 mentions)
Tskgel g4000swxl column, by Tosoh (1 mentions)
Gel filtration standard, by Bio-Rad (1 mentions)
Lc solution software, by Shimadzu (1 mentions)
2 protocols using prominence ultra fast liquid chromatography hplc system
1
SEC Analysis of Monoclonal Antibody
2
Characterization of CRM 197 by SEC-HPLC
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A Shimadzu Prominence ultra fast liquid chromatography HPLC system equipped with a diode array detector (with absorbance detection at 214 nm) was used. Twenty micrograms of each CRM 197 sample was injected onto a TSKgel BioAssist G2SWxl column (7.8 Â 300 mm; Tosoh Biosciences) and the corresponding guard column (Tosoh Biosciences). The columns were operated at 30 C and equilibrated with at least 10 column volumes of mobile phase (10 mM sodium phosphate, 150 mM NaCl, pH 7.2) before sample injection. A flow rate of 0.7 mL/min was used with a 30-min run time. A gel filtration standard (Bio-Rad, Hercules, CA) was subjected to size exclusion chromatography (SEC) before and after the CRM sample to ensure integrity of the column and HPLC system. In addition, the samples were subjected to SEC without the column attached to better determine if insoluble aggregates are present in the samples or if the sample binds to the column (i.e., sample recovery). LC-Solution software (Shimadzu) was used for data analysis.
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