Dados do Trabalho

Name: Next Frontiers to Cure Cancer
Start: 2023-09-15
End: 2023-09-16
Location: Expo Center Norte (Pavilhão Amarelo)

Título

OBTAINMENT AND CHARACTERIZATION OF ANTI-TIM-3 MONOCLONAL ANTIBODY PRODUCED IN BACTERIA

Introdução

Immune checkpoints are negative regulatory receptors on immune cells, maintaining self-tolerance. TIM-3 is implicated in T-cell dysfunction in diseases like cancer and COVID-19. Therapeutic antibodies inhibiting TIM-3 show promise. Bacterial systems offer cost-effective alternatives for antibody production. We investigate anti-TIM-3 antibodies expressed in E. coli.

Objetivo

Our goal is to obtain E. coli-expressed anti-TIM-3 antibodies using phage display. We aim to select functional antibodies, confirm glycosylated epitope interaction, and develop a bacterial system for antibody synthesis, offering an economical approach to therapeutic antibody production.

Métodos

We selected anti-TIM-3 antibodies via phage display, using a human Fab library against TIM-3 ectodomain. Positive clones from 5 rounds of panning were chosen via single-clone ELISA. Clone 4E.Tim3 was expressed in E. coli BL21(DE3) after cloning into pFab. Protein was obtained post-IPTG induction and bacterial lysis. Protein A beads facilitated antibody purification, analyzed using SDS-PAGE. Interaction with glycosylated TIM-3 ectodomain was assessed for functional confirmation.

Resultados

Anti-TIM-3 Fab fragments were successfully expressed in E. coli. Clone 4E.Tim3 showed potential. SDS-PAGE verified purification. Interaction studies suggested glycosylated epitope involvement. This demonstrates bacterial expression's viability for therapeutic antibody production targeting TIM-3.

Conclusões

Innovatively, our study establishes a bacterial system's efficacy in generating functional anti-TIM-3 antibodies. Cost-effectiveness, successful antibody selection, and glycosylated epitope interaction confirmation highlight its potential for large-scale antibody production. This approach contributes to advancing antibody-based therapies for diseases like cancer and COVID-19.