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

被引：15

作者：

Aghamollaei, Hossein ^{[1
]}

Ghanei, Mostafa ^{[1
]}

Rasaee, Mohammad Javad ^{[2
]}

Latifi, Ali Mohammad ^{[3
]}

Bakherad, Hamid ^{[4
,5
]}

Fasihi-Ramandi, Mahdi ^{[6
]}

Taheri, Ramezan Ali ^{[7
]}

Gargari, Seyed Latif Mousavi ^{[8
]}

机构：

[1] Baqiyatallah Univ Med Sci, Chem Injuries Res Ctr, Syst Biol & Poisonings Inst, Tehran, Iran

[2] Tarbiat Modares Univ, Fac Med Sci, Dept Med Biotechnol, Tehran, Iran

[3] Baqiyatallah Univ Med Sci, Appl Biotechnol Res Ctr, Tehran, Iran

[4] Isfahan Univ Med Sci, Dept Pharmaceut Biotechnol, Sch Pharm & Pharmaceut Sci, Esfahan, Iran

[5] Isfahan Univ Med Sci, Isfahan Pharmaceut Sci Res Ctr, Sch Pharm & Pharmaceut Sci, Esfahan, Iran

[6] Baqiyatallah Univ Med Sci, Mol Biol Res Ctr, Tehran, Iran

[7] Baqiyatallah Univ Med Sci, Nanobiotechnol Res Ctr, Tehran, Iran

[8] Shahed Univ, Fac Basic Sci, Dept Biol, Khalij Fars Expressway, Tehran, Iran

来源：

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY | 2021年 / 68卷 / 02期

基金：

美国国家科学基金会;

关键词：

glucose-regulated protein 78; Camelidae antibody; cancer detection; nanobody; unfold protein response; UNFOLDED PROTEIN RESPONSE; ENDOPLASMIC-RETICULUM CHAPERONE; BREAST-CANCER; MONOCLONAL-ANTIBODY; VARIABLE FRAGMENTS; TERMINAL DOMAIN; SURFACE GRP78; VHH NANOBODY; BINDING; IDENTIFICATION;

D O I：

10.1002/bab.1916

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：239 / 246

页数：8

共 44 条

[21] The unfolded protein response: controlling cell fate decisions under ER stress and beyond [J].