Ribosome display and selection of single‐chain variable fragments effectively inhibit growth and progression of microspheres in vitro and in vivo

Abstract

Distinguishing the surface markers of cancer stem cells (CSCs) is a useful method for early diagnosis and treatment of tumors, as CSCs may participate in tumorigenesis and metastasis by migrating into the circulatory system. However, the potential targets of CSCs are expressed at low levels in the natural state and are always changing. Thus, dynamic screening has been reported to be an effective measure for exploring CSC markers. In recent years, diverse single‐chain variable fragments (scFvs) have been widely used in immunotherapy. In this study, we determined that the scFvs, screened using RD, had a high affinity to microspheres and could inhibit their progression. We also observed that the selected scFvs underwent evolution in vitro, and antitumor‐associated proteins were successfully expressed. Combined with chemotherapy, the scFvs had a synergistic effect on the inhibition of the microspheres’ progression in vitro and in vivo, which could be ascribed to their high affinity for stem‐like cells and the inhibition of the microspheres’ collective behaviors. In addition, proteins inhibiting CD44+/CD24+ and MAPK were involved. Our data indicated that dynamic screening of the scFvs in a natural state was of great significance in the inhibition of the microspheres in vitro and in vivo.

CD24: A Novel Target for Cancer Immunotherapy

“…Recombinant antibodies targeting CD24 have also been tested. Single-chain variable fragments (scFvs), selected from a total RNA library from lymphocytes of breast cancer patients, showed high selectivity for CD24 and CD44 and produced a synergistic effect when administered in combination with epirubicin [ 109 ]. ScFvs have received FDA approval in malignancies, including blinatumomab for acute lymphoblastic leukemia [ 119 ], and represent an alternative method for targeting CD24.…”

Rapid-format recombinant antibody-based methods for the diagnosis of Clostridioides difficile infection: Recent advances and perspectives

“…However, one of the limitations of this method is the accessible, functional ribosome levels in the reaction for the library, which relates to the library size ( Kunamneni et al, 2020 ). The ribosome display technology has been applied for selecting scFvs with high affinity against different targets such as tumor cells ( Huang et al, 2018 ), Zika virus (ZIKV) ( Kunamneni et al, 2018 ), Mycobacterium tuberculosis ( Ahangarzadeh et al, 2017 ), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ( Chen et al, 2021 ). Moreover, it has been successfully used for scaffold selection, including affibodies ( Lagoutte et al, 2019 ), however, there are no reports about antibody selection through ribosome display for CDI diagnosis or treatment.…”

Ribosome Display Technology: Applications in Disease Diagnosis and Control

“…Ribosome display is a powerful and ideal in-vitro tool that can perform such screening tasks for highly specific and high-affinity single-chain antibodies [ 54 , 112 ]. Reasoning along this line, Huang et al [ 113 ] applied ribosome display to perform large-scale screenings of scFvs against tumor cells. scFvs with high affinity for cancer stem cells were obtained in their study, and the activities of these scFvs could hinder the growth of cancer cells in vitro and in vivo.…”