Bioanalytical

  • SEC analysis of Bevacizumab (Avastin®) and its aggregates and fragments

SEC analysis of Bevacizumab (Avastin®) and its aggregates and fragments

Jun 24 2020 Read 428 Times

Due to the sizes of monoclonal antibodies (MAbs, about 150 kDa), size exclusion chromatography (SEC) is a standard technique for analysing MAbs such as Bevacizumab (Avastin®). It is also a standard separation mode used in quality control to obtain information about aggregation and/or fragmentation of the MAbs.

In this application note YMC’s dedicated SEC column for antibodies, YMC-SEC MAB, is used to separate the monomer antibody from its aggregates as well as from degradation products in a single run. A neutral phosphate buffer containing 0.2 M NaCl is used as eluent at a flow rate of 0.165 mL/min. UV detection is carried out at 280 nm. The resulting separation provides high resolutions for both the high molecular weight range as well as for the low molecular weight range. The resulting backpressure is only 34 bar/3.4 MPa, while the maximum pressure rating for the column is 140 bar/14 MPa.

YMC-SEC MAB is a column specially designed for the simultaneous analysis of antibodies, their fragments or aggregates. The column is a silica-based size exclusion chromatography column utilising a dihydroxypropyl stationary phase bonded to a silica gel base, specifically designed for antibody separations.

This material offers a 250 Å pore size in 3 µm particle size, making it an excellent choice for separating aggregates from monomer, as well as effectively resolving lower molecular weight fragments. YMC-SEC MAB exhibits excellent resolution and peak shape over a wide molecular weight range of 10 kDa to 700 kDa. Of course, YMC-SEC MAB ensures excellent lot-to-lot reproducibility, which is ideal for quality control.

Reader comments

Do you like or dislike what you have read? Why not post a comment to tell others / the manufacturer and our Editor what you think. To leave comments please complete the form below. Providing the content is approved, your comment will be on screen in less than 24 hours. Leaving comments on product information and articles can assist with future editorial and article content. Post questions, thoughts or simply whether you like the content.

Post a Comment





Digital Edition

Chromatography Today - June 2020

June 2020

In This Edition Articles Fundamental Aspects - Separation of the 4 Enantiomers of the Fungicide Spiroxamine by LC-MS/MS - Optimising the Chiral Separation of the Pesticide Diniconazole...

View all digital editions

Events

BMSS Annual Meeting - CANCELLED

Sep 08 2020 Sheffield, UK

Mass Spec 2020

Sep 14 2020 Virtual Symposium

ISC 2020 - POSTPONED TO 2022#

Sep 21 2020 Budapest, Hungary

MSB 2020 - NEW DATES

Sep 27 2020 Saint-Malo, France

CE Pharm 2020 - ONLINE EVENT

Sep 28 2020 Bethesda, MD, USA

View all events