HPLC Analysis of Reduced SILu™ Lite SigmaMAb on Zenix® SEC-300 with pH-Neutral Buffer

HPLC Analysis of Reduced SILu™ Lite SigmaMAb on Zenix® SEC-300 with pH-Neutral Buffer

Conditions

column Zenix SEC-300, 30 cm x 4.6 mm I.D., 3 µm particles (Z777028)
column temp. 25 °C
mobile phase [A] 0.2 M sodium phosphate dibasic, pH 7.0 with phosphoric acid; [B] water; [C] acetonitrile; (50:20:30, A:B:C)
flow rate 0.3 mL/min
pressure 1070 psi (74 bar)
sample reduced SigmaMAb (with 10 mM dithiobutylamine), 1 g/L, in mobile phase A
injection 1 µL
detector UV, 215 nm

Description

Analysis Note The integrity of antibodies in the development of biologics is of the utmost importance in the pharmaceutical quality control lab; therefore, a method to examine intact antibodies versus reduced antibody subunits is needed. Separation of the light and heavy chains of the SigmaMAb antibody standard, was achieved using size exclusion chromatography on a Zenix SEC-300 column.
Categories Analytical Chromatography, Antibodies, Proteins
Featured Industry Life Science and Biopharma
Legal Information SILu is a trademark of Sigma-Aldrich Co. LLC, Zenix is a registered trademark of Sepax Technologies
suitability application for HPLC

Materials

     
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