SEC-HPLC and cation-exchange chromatography (CEX-HPLC) were employed to analyse the thermostability of trivalent IL-5-HSA Nb. Briefly, the trivalent IL-5-HSA Nb was diluted to 1 mg/mL and incubated at 2–8 °C or 25 °C for 1 month or even under 3 freeze-and-thaw cycles. After incubation, the stability of Nb was measured by SEC-HPLC and CEX-HPLC analyses. CEX-HPLC was performed on a Dionex Ultimate 3000 RSLC system (Thermo Fisher Scientific) with a BioMab NP5 PK column (Agilent Technologies).
Biomab np5 pk column
Manufactured by Agilent Technologies
The BioMab NP5 PK column is a high-performance liquid chromatography (HPLC) column designed for the analysis of monoclonal antibodies (mAbs) and other biopharmaceuticals. The column features a non-porous, silica-based stationary phase that provides efficient separation and high recovery of target molecules. It is intended for use in the characterization of protein therapeutics, including pharmacokinetic (PK) studies.
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Lab products found in correlation
2 protocols using biomab np5 pk column
1
Stability analysis of trivalent IL-5-HSA Nb
2
Charge Variant Quantification of BioMab
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Weak cation-exchange chromatography (WCX) was performed to quantify
charge variants on a BioMab, NP5, PK column (4.6 mm × 250 mm,
5 μm, Agilent) operated at 25 °C using a Dionex Ultimate
3000 RSLC system (Thermo Scientific). Prior to injection, the column
was saturated with 65% mobile phase A (15 mM sodium phosphate buffer
and 0.05% NaN3 at pH 6.2) and 35% mobile phase B (150 mM
sodium phosphate buffer and 0.05% NaN3 at pH 6.2). All
buffers were filtered with 0.22 μm filters. Charged species
were separated using a runtime of 23 min with a linear gradient from
35 to 65% B at a flow rate of 0.800 mL/min. The samples were diluted,
depending on UV-based concentrations after each cycle (dilution factor
∼4 if concentration ∼20 mg/mL), to make final concentration
of 5 mg/mL for each sample, and 30 μg of protein was injected
(in triplicates) into the column.
charge variants on a BioMab, NP5, PK column (4.6 mm × 250 mm,
5 μm, Agilent) operated at 25 °C using a Dionex Ultimate
3000 RSLC system (Thermo Scientific). Prior to injection, the column
was saturated with 65% mobile phase A (15 mM sodium phosphate buffer
and 0.05% NaN3 at pH 6.2) and 35% mobile phase B (150 mM
sodium phosphate buffer and 0.05% NaN3 at pH 6.2). All
buffers were filtered with 0.22 μm filters. Charged species
were separated using a runtime of 23 min with a linear gradient from
35 to 65% B at a flow rate of 0.800 mL/min. The samples were diluted,
depending on UV-based concentrations after each cycle (dilution factor
∼4 if concentration ∼20 mg/mL), to make final concentration
of 5 mg/mL for each sample, and 30 μg of protein was injected
(in triplicates) into the column.
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