Repository of Research and Investigative Information

Repository of Research and Investigative Information

Baqiyatallah University of Medical Sciences

Isolation and characterization of a novel nanobody for detection of GRP78 expressing cancer cells

(2021) Isolation and characterization of a novel nanobody for detection of GRP78 expressing cancer cells. Biotechnol Appl Biochem. pp. 239-246. ISSN 0885-4513

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Abstract

Glucose-regulated protein 78 (GRP78) is an endoplasmic reticulum (ER) chaperone that has been shown that is overexpressed in cancer cells. Overexpression of GRP78 on cancer cells makes this molecule a suitable candidate for cancer detection and targeted therapy. VHH is the binding fragment of camelid heavy-chain antibodies also known as "nanobody." The aim of this study is to isolate and produce a new recombinant nanobody using phage display technique to detect cancer cells. Using the c-terminal domain of GRP78 (CGRP) as an antigen, four rounds of biopanning were performed, and high-affinity binders were selected by ELISA. Their affinity and functionality were characterized by surface plasmon resonance (SPR) cell ELISA and immunocytochemistry. A unique nanobody named V80 was purified. ELISA and SPR showed that this antibody had high specificity and affinity to the GRP78. Immunofluorescence analysis showed that V80 could specifically bind to the HepG2 and A549 cancer cell lines. This novel recombinant nanobody could bind to the cell surface of different cancer cells. After further evaluation, this nanobody can be used as a new tool for cancer detection and tumor therapy.

Item Type: Article
Keywords: A549 Cells Antineoplastic Agents, Immunological/*immunology Endoplasmic Reticulum Chaperone BiP Gene Expression Regulation, Neoplastic/*immunology Heat-Shock Proteins/genetics/*immunology Hep G2 Cells Humans Neoplasm Proteins/genetics/*immunology Neoplasms/genetics/*immunology/pathology Single-Domain Antibodies/genetics/*immunology Camelidae antibody cancer detection glucose-regulated protein 78 nanobody unfold protein response
Page Range: pp. 239-246
Journal or Publication Title: Biotechnol Appl Biochem
Journal Index: Pubmed
Volume: 68
Number: 2
Identification Number: https://doi.org/10.1002/bab.1916
ISSN: 0885-4513
Depositing User: مهندس مهدی شریفی
URI: http://eprints.bmsu.ac.ir/id/eprint/9428

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