Proteomics-Based Platform for Monoclonal Antibody Preparation May Replace Hybridoma Techniques

(Danvers, MA, USA) has announced the release of its NG-XMT monoclonal antibody discovery platform. The methodology underlying this platform was described in a paper published in the May 2012 issue of the journal Nature Biotechnology.

The NG-XMT system is based on Cell Signaling Technology’s PTMScan proteomics technology, which employs proprietary techniques for antibody-based peptide enrichment combined with tandem mass spectrometry for quantitative profiling of post-translational modifications (PTMs), including phosphorylation, ubiquitination, and acetylation.

The new approach involves affinity purification of antibodies with high specific activity and then analyzing digested antibody fractions by nano-flow liquid chromatography coupled to tandem mass spectrometry.

High-confidence peptide spectral matches of antibody variable regions were obtained by searching a reference database created by next-generation DNA sequencing of the B-cell immunoglobulin repertoire of the immunized animal. Finally, heavy and light chain sequences were paired and expressed as recombinant monoclonal antibodies. Using this technology, investigators at Cell Signaling Technology isolated monoclonal antibodies for five antigens from the sera of immunized rabbits and mice. The antigen-specific activities of the monoclonal antibodies recapitulated or surpassed those of the original affinity-purified polyclonal antibodies.

“Our NG-XMT proteomics-based technology is faster and more efficient than traditional hybridoma-based methods,” said senior author Dr. Roberto D. Polakiewicz, CSO of Cell Signaling Technology. “Moreover, our proprietary and patent pending technology can be applied to the development of therapeutically relevant human monoclonal antibodies for vaccine development and for understanding basic questions in immunology. Cell Signaling Technology has long been an industry leader in the development and manufacture of high performance antibodies for basic and clinical research, and we are excited to announce this breakthrough in monoclonal antibody technology.”

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