Construction of a camel-derived natural phage nanobody display library and screening of anti-CD22 nanobodies.
- Author:
Wanjun HE
1
;
Kai CUI
1
;
Xiqian ZHANG
2
;
Dan JIANG
1
;
Guangxian XU
3
Author Information
1. School of Medical Technology, Guangdong Medical University, Precision Medicine Center affiliated to Dongguan First Hospital, Guangdong Medical University, Guangdong Provincial Key Laboratory of Medical of Medical Immunology and Molecular Diagnostics, Guangdong Medical University, Dongguan 523000, China.
2. Guangdong Provincial Key Laboratory of Medical of Medical Immunology and Molecular Diagnostics, Guangdong Medical University, Dongguan 523000, China.
3. School of Medical Technology, Guangdong Medical University, Precision Medicine Center affiliated to Dongguan First Hospital, Guangdong Medical University, Guangdong Provincial Key Laboratory of Medical of Medical Immunology and Molecular Diagnostics, Guangdong Medical University, Dongguan 523000, China. *Corresponding author, E-mail: xuguangxian@gdmu.edu.cn.
- Publication Type:Journal Article
- MeSH:
Camelus/immunology*;
Single-Domain Antibodies/chemistry*;
Peptide Library;
Humans;
Animals;
Sialic Acid Binding Ig-like Lectin 2/genetics*;
Amino Acid Sequence;
Molecular Docking Simulation
- From:
Chinese Journal of Cellular and Molecular Immunology
2025;41(3):254-261
- CountryChina
- Language:Chinese
-
Abstract:
Objective To screen the anti-CD22-specific nanobodies to provide a basis for immunotherapy agents. Methods The naive phage nanobody library was constructed and its diversity was analyzed. Three rounds of biotinylated streptavidin liquid phase screening were performed by using biotinylated CD22 antigen as the target, and the sequence of nanobodies against CD22 were identified by ELISA and gene sequencing. Results The capacity of the constructed naive phage nanobody library was 3.89×109 CFU/mL, and the insertion of effective fragments was higher than 85%. Based on this library, seven anti-human CD22 nanobodies were screened, and the amino acid sequence comparison results showed that the overall similarity was 70.34%, and all of them were hydrophilic proteins. The results of protein-protein complex docking prediction showed that the mimetic proteins of the five nanobody sequences could be paired and linked to CD22, and the main forces were hydrophobic interaction and hydrogen bonding. Conclusion This study provided a basis for the study of chimeric antigen receptor T cells targeting CD22, successfully constructed the natural phage nanobody library and obtaining five anti-CD22-specific nanobodies.
