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A blog on the latest applications, articles, & research on chromatography solutions in sample preparation, Ion Chromatography (IC),
High Pressure Liquid Chromatography (HPLC), Ion Chromatography-Mass Spectrometry (IC-MS), Gas Chromatography (GC),
Gas Chromatography-Mass Spectrometry (GC-MS and GC-MS/MS), and software (Chromeleon CDS, LIMS, and ProteinCenter).

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HPLC Column Improves Separation of a Monoclonal Antibody


Monoclonal antibodies analysisRecently, an interesting study on the clinical development of monoclonal antibody-based drugs in HIV and HCV diseases (link to full, free article) crossed my desk that made for interesting reading as it discussed the advantages in using monoclonal antibodies (MAbs) in the treatment of infectious diseases, such as HIV and HCV, as more viruses become resistant to current drugs. The advantages of MAbs include low toxicity, highly specific interactions, good biological compatibility, and high versatility.

(For those who are new to MAbs and their potential, check out this American Cancer Society webpage on the use of MAbs in the treatment of cancer.)

Typically, MAbs are analyzed both in the intact form and after cleavage using ion-exchange chromatography and size-exclusion chromatography but here we present a novel, reversed-phase HPLC separation technique that is compatible with mass spectrometry. 

The method uses one of our newer solid-core HPLC columns (Thermo Scientific Accucore 150-C4 HPLC column) to provide a fast, high-efficiency separation of the entire antibody as well as the fragmented region along with low-level impurities resulting from fractionation of the MAb. The researcher can then introduce the intact proteins to high-resolution mass spectrometry for accurate molecular weight determination and identification of variants.

The method is described in Application Note 20570, Reversed-Phase Analysis of a Monoclonal Antibody (MAb) on an Accucore 150-C4 HPLC Column, (downloadable PDF) takes 20 mins to complete at pressures compatible with conventional HPLC instrumentation for the analysis of a monoclonal antibody used to treat chronic lymphocytic leukemia.

The method was developed on our biocompatible HPLC system (Thermo Scientific Dionex UltiMate U3000 Titanium system) equipped with an autosampler (Thermo Scientific Dionex WPS-3000PL autosampler) and a variable wavelength detector (Thermo Scientific Dionex VWD3400 detector). Data analysis and instrumentation control was managed using our Chromatography Data System (CDS) software (Thermo Scientific Dionex Chromeleon 6.8 SR11 CDS software).

This blog has over 25 published blog posts on MAb applications, webinars, market trends, and more that you can also check out for more information.

Also, don't forget to add your questions/comments to the Comments box below; our experts will be pleased to hear from you.


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