<p>Preparation and Characterization of Anti-GPC3 Nanobody Against Hepatocellular Carcinoma</p>

Abstract

Background: Glypican-3 (GPC3) is a newly identified target molecule for the early diagnosis of hepatocellular carcinoma (HCC), while targeted inhibition of GPC3 signaling may help to control the proliferation and metastasis of HCC cells. The purpose of this study was to prepare the anti-GPC3 nanobody and to investigate the affinity of the anti-GPC3 nanobodies in vitro and the anticancer effects on hepatocellular carcinoma in vivo. Methods: To screen for unknown anti-GPC3 antibodies, we constructed an antibody phage display library. After three rounds of panning, positive phage clones were identified by enzyme-linked immunosorbent assay (ELISA). Further, the nanobody fusion protein was expressed in E. coli BL21 cells and purified by affinity chromatography. Competitive ELISA and flow cytometry were conducted to confirm the affinity of the anti-GPC3 nanobodies in vitro. The antitumor effects of VHH GPC3 were assessed in vivo. Results: The results showed that the nanobody VHH GPC3 had specific high-affinity binding to His-GPC3 antigen. Moreover, VHH GPC3 exhibited specific binding to commercial human GPC3 and recognized the surface GPC3 protein of the hepatoma cell line HepG2. Importantly, in vivo study showed that GPC3 nanobody suppresses the growth of HepG2 and improves the survival rate of tumor mice. Discussion: In summary, our new anti-GPC3 nanobody suggests a strong application potential for targeted therapy of liver cancer.

Prospect of SARS-CoV-2 spike protein: Potential role in vaccine and therapeutic development

“…This study also suggests that lipidation, as well as linker length, plays an essential role in the overall activity of lipopeptides. (S. L. Xia et al, 2020 ; S. Xia et al, 2020 ). This would be a potential candidate for therapeutic development.…”

“…For example, nanobodies inhibiting PD1-PDL1 interaction, LIN28:let-7 interaction, and T cell marker CD3 are all promising candidates for targeting and activating cytotoxic T lymphocytes (CTLs) to induce anti-tumor responses. GRP78, CD38, and GPC3 expression on cancer cells have also been targeted for nanobody expression and purification, which provide useful cancer cell markers and potential therapeutic markers ( Aghamollaei et al, 2020 , p. 78; Hambach et al, 2020 ; Li et al, 2020 ; Moradi-Kalbolandi et al, 2020 ; L. L. Xia et al, 2020 ; S. Xia et al, 2020 ; C. J. Yu et al, 2020 ; C. Yu et al, 2020 ).…”

Preparation of a Nanobody Specific to Dectin 1 and Its Anti-inflammatory Effects on Fungal Keratitis

“…A total of 95 phages were randomly picked from the third round panning and then analyzed by monoclonal phage ELISA, and 26 showed high absorbance (at least three times that of control wells, Figure 1C). 25 Four phages with the highest absorbance were selected to repeat the phage ELISA in order to avoid false-positive reactivity. The absorbance of these was at least three times that of the negative control (Figure 1B).…”

The transcatheter arterial chemoembolization combined with targeted nanoparticle delivering sorafenib system for the treatment of microvascular invasion of hepatocellular carcinoma

“…GPC3 can become the target molecule of HCC, and it can also induce the ADCC response to play an antitumor role by using the blocking antibody HGC33 [ 29 ]. As a glycoprotein that can penetrate cell membranes, GPC3 had a specific expression of nearly 80% for HCC [ 30 , 31 ]. Yao et al (2013) [ 32 ] used immunohistochemical method to detect GPC3 in both HCC and non-hepatocellular tumors.…”