Antibody Sequencing and Quantitation

Theo Luider
Erasmus MC, Rotterdam, the Netherlands

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Theo Marten Luider (Presenter) Erasmus MC

Presenter Bio: The major aim of our research group is to investigate various diseases such as glioma, multiple sclerosis and Alzheimer by mass spectrometric approaches. In addition, we collaborate on various topics as broad as breast cancer, prostate cancer, colorectal cancer, lung carcinoma and topics related to virology and medical microbiology. Expertise is present on identification of proteins in complex samples as well as quantification of specific proteins in complex samples. We perform laser microdissection technology and have developed various techniques to get informative mass spectrometry from minute numbers of cells ( lower than 1000 cells). The technical possibility of combining state of the art mass spectrometry with other omics technologies opens ways to answer essential medical questions related to pathways involved in cancer and discovery of antibodies.

Relevant Financial Disclosures (within past 24 months) Discussing Products? NO, presenter will NOT present educational content related to products/services of the ineligible company(ies) listed above. Conflict mitigation NOT REQUIRED.

The metabotropic glutamate receptor-1 (mGluR1) has recently been identified as an oncogene that is abundantly expressed in >60% of human melanoma, breast, renal cell and prostate cancer tissues. Many of these cancers cannot be successfully treated and new treatment modalities are clearly needed. In 2003, glutamate signalling through mGluR1 was reported to be a promising new molecular target for the treatment of cancer. We identified that affinity purified mGluR1 autoantibodies from plasma from a unique ataxia patient effectively block the mGluR1 receptor both in vitro and in vivo. These fully matured human antibodies are selective and have high target affinity. The precise sequence of these antibodies would offer an attractive basis for the development of a new antibody-based therapy for mGluR1 dependent cancers.

We have developed a novel technology pipeline for mass spectrometry (MS) based protein sequencing of full length human antibodies from plasma and reveal the full length sequence of the anti-mGLuR1 antibodies. The recovery of fully matured human derived autoantibody sequences with this pipeline has great potential for therapeutics. Existing antibody-based therapies rely on antibodies raised in other species, which can evoke anti-drug responses and are less efficient than human autoantibodies. In order to successfully develop the MS based protein sequencing pipeline, we have optimized antibody enrichment from plasma samples, advance novel protocols for MS sample preparation, and develop state-of-the-art MS analysis tools to derive protein sequences from the complex data output. In this way, we will produce an anti-mGluR1 antibody with therapeutic potential, and a pipeline that allows for the recovery of additional fully matured human derived antibody sequences for other indications like multiple myeloma.