Trường Đại học Khoa Học Tự nhiên

Cơ sở 1: 227 Nguyễn Văn Cừ, Phường 4, Quận 5, TP Hồ Chí Minh

Cơ sở 2: Khu phố 6, Phường Linh Trung, TP Thủ Đức, TP Hồ Chí Minh

Screening of materials-binding peptides and application to site-directed immobilization of antibody fragments

14:25, 09/10/2023
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Screening of materials-binding peptides and application to site-directed immobilization of antibody fragments

Dr. Yoichi Kumada, Faculty of Molecular Chemistry and Engineering, Kyoto Institute of Technology

 

Abstract
Immobilization and orientation control of functional proteins on the surfaces of materials has been regarded as a key factor for enhancement of performance of protein-based materials in the fields of bio-production, diagnostics and clinical treatments. Especially, Sensitive and accurate biomolecular sensing utilizing antibody-immobilized solid support of the solid-sensing materials are highly expected in clinical diagnosis, pharmaceutical process monitoring and environmental analysis. Here, we propose and demonstrate development of materials-binding peptides for site-specific immobilization of recombinant antibody fragments, such as single-chain Fv antibodies and single-domain antibodies (VHH).

We have successfully identified polystyrene-binding peptides (PS-tag) as well as poly(methylmethacrylate)-binding peptide (PMMA-tag) and the others from the original peptide/protein library stocks utilizing proteomic analysis technologies. Genetic fusion of materials-binding peptides at the C-terminal regions of recombinant antibody fragments resulted in enhancement of antigen-binding signals that were detected from the plates, indicating increase in both density and residual activity of antibody fragments on the solid surface. We further applied our technologies to SPR sensor that the sensing area was coated with thin layer of plastics. Both PS-tag-fused scFv and PMMA-tag-fused scFv were preferentially attached on these plastic surfaces at the higher density, and consequently, much higher antigen-binding signals were detected. These results clearly indicated that combination of materials-binding peptides and plastic thin nano-layer would be a promising interface to immobilize recombinant antibody fragments site-specifically.
 

Poster_Kumada-sensei

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