Agilent HPLC-Chip 6520 Accurate-Mass LC/MS Application Note

This application note describes the characterization of a monoclonal antibody at low nanogram levels using an Agilent HPLC-Chip system coupled to an Agilent Accurate-Mass Q-TOF MS instrument. The superior sensitivity and mass accuracy of the HPLC-Chip LC/MS platform, combined with the powerful data processing capabilities of Agilent MassHunter and BioConﬁrm software, enabled easy and rapid identiﬁ cation of antibody heterogeneity and cleaved fragments. Robust, reliable, and easy to use, Agilent nanoﬂ ow LC/MS technology is ideally suited for the routine analysis of biopharmaceuticals.

Brand: Agilent
Document type: Application Note
Language: English
Pages: 12
File format: PDF
File size: 1.1 MB
Published: 02 July, 2012
Updated: 30 September, 2023
MD5 checksum: C2392DE2466A496858D18AA189E3D617
PDF: Agilent HPLC-Chip 6520 Accurate-Mass LC/MS Application Note PDF

About this manual

The document details the analysis of a recombinant monoclonal antibody (mAb) at low nanogram levels. It utilizes an Agilent HPLC-Chip system coupled with an Agilent 6520 Accurate-Mass Q-TOF LC/MS platform to identify antibody heterogeneity and cleaved fragments.

What's inside

Introduction
Experimental Materials
Deglycosylation Reaction
Papain Digestion
Reduction Reaction
LC/MS Analysis Instrumentation
Data Analysis Software

Frequently asked questions

What is the flow rate for the enrichment column?

The flow rate from the capillary pump to the enrichment column is 3 pL/min.

What is the minimum amount of intact monoclonal antibody required for on-column analysis?

The procedure requires a sample load of 7.5 to 10 ng on-column for intact mAb.

What is the incubation temperature and duration for the papain digestion step?

The mixture must be incubated at 37°C for 2 hours.

What is the measured mass accuracy obtained for the heavy chain after reduction?

The mass accuracy obtained is 1.9 ppm.

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