Anti-tetanus human monoclonal antibodies.

Author(s)

Eduardo Aliprandini

• Document type: Doctoral Thesis
• Press: São Paulo.
• Institution: Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI)
• Defense date: 2015-08-12

Examining board members

Ana Maria Moro; Silvia Beatriz Boscardin; Ana Paula Lepique; Daniela Santoro Rosa; Marco Antonio Stephano

• Advisor: Ana Maria Moro; Silvia Beatriz Boscardin

Abstract

Monoclonal antibodies (mAbs) for therapeutic use correspond to a major area of the biopharmaceutical industry, especially human mAbs that are less prone to elicit immunogenicity. The objective of this work was to obtain anti-tetanus human mAbs through separation of memory B lymphocytes producing specific antibodies stained with the antigen or plasmablasts. Cells were collected from peripheral blood of donors after vaccination and separated through cell sorting. The variable regions of the antibodies were amplified and cloned in expression vectors for transient transfection of HEK293-F cells. The staining with the tetanus toxin labeled independently with two markers, biotin and Alexa Fluor® 647 allowed the separation of specific B lymphocytes producing anti-tetanus mAbs. The antibodies expressed were evaluated by ELISA, western blotting and the inhibition of the binding of the tetanus toxin to the ganglioside GT1b. The in vivo neutralization assay showed that a pool of three different mAbs were able to protect mice against the tetanus toxin. (AU)

• FAPESP's process: 11/22334-2 - Human Monoclonal Antibodies Anti-Tetanus Toxoid
• Grantee: Eduardo Aliprandini
• Support Opportunities: Scholarships in Brazil - Doctorate