Analytical HIC-HPLC for non-destructive mAb characterization

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Monitor Post-Translational Modifications

with Sepax Proteomix® HIC-Butyl 5µm

  • Using a simple native condition without IPA additives, Sepax Proteomix HIC is able to provide the oxidative variant separation based on sample hydrophobicity. The variants generated were on the industry standard NIST mAb. Peaks eluted prior to the main peak could possibly be the oxidation of the antibody or structural changes by aspartic acid isomerization.[1]
  • Sepax Proteomix® HIC-Butyl 5µm is able to provide an orthogonal method to MS for mAb characterization under native conditions
  • Proteomix® HIC 5µm and 10µm is also available in column IDs up to 7.5 cm preparative sizes for lab scale purification and fractionation.
  • The same particles allow for complete linear scalability from analytical to preparative.
NIST MAb on Proteomix HIC Butyl-NP5

Column: Proteomix HIC Butyl-NP5, 5um, NP 4.6 x 100 mm (PN:431NP5-4610); Mobile Phase: A: 2 M Ammonium Sulfate, in 100 mM Sodium Phosphate pH 7.0, B: 100 mM Sodium Phosphate pH 7.0; Flow rate: 0.5 mL/min; System: HPLC; Column Temperature: 30 ℃; Detection: UV 280 nm; Sample: NIST MAb 10 mg/mL (pI 9.18, in 12.5 mM histidine, pH 6.0);  Injection volume: 2 µL

 [1] Separation Methods and Orthogonal Techniques
David A. Michels, Anna Y. Ip, Thomas M. Dillon, Kurt Brorson, Scott Lute, Brittany Chavez, Ken M. Prentice, Lowell J. Brady, and Karen J. Miller
State-of-the-Art and Emerging Technologies for Therapeutic Monoclonal Antibody Characterization Volume 2. Biopharmaceutical Characterization: The NISTmAb Case Study. January 1, 2015 , 237-284
DOI:10.1021/bk-2015-1201.ch005

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Proteomix® HIC-Butyl is a non-porous media applicable for analytical analysis, laboratory discovery, laboratory-scale purification and preparative chromatography for the production of several milligrams to grams of proteins. Sepax also provides polymethacrylate-based Generik MC and Polar MC HIC 30 µm and 60 µm media used for polishing and/or industrial scale purification.

Other HIC Application Data:

 
GiMiTEC ID# 1376300
GiMiTEC CPL# https://channel.gimitec.com/node/1376300

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