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Biochromatography - Intact and Fragmented Protein Profiling

The analysis of intact and fragmented proteins is an essential step in the characteristation of a protein.  This can be used to identify and monitor potential degradation products throughout development, manufacturing and storage. 

Reversed-phase chromatography is an excellent way of analysing the intact and reduced proteins including light and heavy chain (Lc/Hc), fragment crystallisable and fragment antigen binding (Fc/Fab) and other post translation modifications.

One of the benefits of using reversed-phase is that the methods are generally compatible with MS detection which allows accurate mass determination.

Product Information

Crawford Scientific can help you separate and characterise biomolecules with our family of reversed phase BioHPLC columns.

Due to their size proteins diffuse very slowly through a column. Faster mass transfer can be acheived by using short chain length ligands (C3 or C4), large pore size, small particles and superfically porous particles. For more detailed product information click on the links below:

Agilent AdvanceBio RP-mAb columns

Agilent Poroshell 300Å columns

Agilent Zorbax 300Å Stablebond columns

Agilent Zorbax 300Å Extend columns

ThermoScientific Accucore 150Å columns

ThermoScientific ProSwift RP-10R

Thermo Scientific SMART Digest™ Kits



Please contact us to discuss your specific biochromatography needs

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Application notes

Here at Crawford Scientific we have gathered all of the most relevant and useful documentation, including application notes and some general guides, relating to protein characterisation using reversed-phase chromatography, see below;


Protein and Impurity Profiling Using Wide Pore Reversed Phase HPLC

Reversed Phase Characterisation of Biomolecules with Advancebio Columns

Separation of High Molecular Weight Fibrous Proteins

Using Temperature to Increase Resolution in Ultrafast Protein Separations

Poroshell300 Fast Protein Digest Analysis

Disulfide Linkage Analysis of IgG1 using an Agilent 1260 Infinity Bio-inert LC System with an Agilent ZORBAX RRHD Diphenyl sub-2 μm Column

Reversed-Phase Separation of Intact Monoclonal Antibodies Using Agilent ZORBAX Rapid Resolution High Definition 300SB-C8 1.8 μm Column

Reversed-Phase Optimization for Ultra-Fast Profiling of Intact and Reduced Monoclonal Antibodies Using ZORBAX Rapid Resolution High Definition 300SB-C3 Column

N-Terminal Site-Specific PEGylation and Analytical-Scale Purification of PEG Lysozyme

Ultra High Speed and High Resolution Separations of Reduced and Intact Monoclonal Antibodies with Agilent ZORBAX RRHD Sub-2 μm 300 Diphenyl UHPLC Column

Fast Separation of Recombinant Human Erythropoietin Using Reversed Phased Agilent ZORBAX RRHD 300SB-C18, 1.8 μm


Reversed-Phase Analysis of a Monoclonal Antibody on an Accucore 150-C4 HPLC Column



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