Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Marie-Claire Devilder 1 Melinda Moyon 1 Xavier Saulquin 1, * Laetitia Gautreau-Rolland 1, *
* Corresponding author
1 CRCINA - Département INCIT - Equipe 1 - Immunobiology of Human αβ and γδ T cells and Immunotherapeutic Applications
CRCINA - Centre de recherche de Cancérologie et d'Immunologie / Nantes - Angers
Abstract : Monoclonal antibodies (mAbs) are powerful tools useful for both fundamental research and in biomedicine. Their high specificity is indispensable when the antibody needs to distinguish between highly related structures (e.g., a normal protein and a mutated version thereof). The current way of generating such discriminative mAbs involves extensive screening of multiple Ab-producing B cells, which is both costly and time consuming. We propose here a rapid and cost-effective method for the generation of discriminative, fully human mAbs starting from human blood circulating B lymphocytes. The originality of this strategy is due to the selection of specific antigen binding B cells combined with the counter-selection of all other cells, using readily available Peripheral Blood Mononuclear Cells (PBMC). Once specific B cells are isolated, cDNA (complementary deoxyribonucleic acid) sequences coding for the corresponding mAb are obtained using single cell Reverse Transcription-Polymerase Chain Reaction (RT-PCR) technology and subsequently expressed in human cells. Within as little as 1 month, it is possible to produce milligrams of highly discriminative human mAbs directed against virtually any desired antigen naturally detected by the B cell repertoire.
Document type :
Journal articles
Complete list of metadatas

Cited literature [13 references]  Display  Hide  Download

https://www.hal.inserm.fr/inserm-01745629
Contributor : Elizabeth Bernardo <>
Submitted on : Wednesday, March 28, 2018 - 1:54:30 PM
Last modification on : Monday, May 20, 2019 - 3:32:01 PM

File

 Restricted access
To satisfy the distribution rights of the publisher, the document is embargoed until : jamais

Please log in to resquest access to the document

Identifiers

Collections

Citation

Marie-Claire Devilder, Melinda Moyon, Xavier Saulquin, Laetitia Gautreau-Rolland. Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood. Journal of visualized experiments : JoVE, JoVE, 2018, pp.e56508. ⟨10.3791/56508⟩. ⟨inserm-01745629⟩

Share

Metrics

Record views

158