Generation of mWasabi fluorescent protein-binding nanobodies

Shufeng Li; Haitao Shan; Ting Wang; Xiaojuan Zheng; Minke Shi; Baojun Chen; Haiqin Lu; Ying Zhang; Sheng Zhao; Zichun Hua

doi:10.1016/j.ab.2020.113875

Generation of mWasabi fluorescent protein-binding nanobodies

Anal Biochem. 2020 Nov 1:608:113875. doi: 10.1016/j.ab.2020.113875. Epub 2020 Jul 31.

Authors

Shufeng Li¹, Haitao Shan², Ting Wang², Xiaojuan Zheng², Minke Shi³, Baojun Chen³, Haiqin Lu², Ying Zhang², Sheng Zhao², Zichun Hua⁴

Affiliations

¹ The Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Southeast University Medical School, 87 Dingjiaqiao Road, Nanjing, 210009, China. Electronic address: shufengli@seu.edu.cn.
² The Laboratory of Developmental Genes and Human Diseases, Ministry of Education, Southeast University Medical School, 87 Dingjiaqiao Road, Nanjing, 210009, China.
³ Department of Thoracic and Cardiovascular Surgery, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, China.
⁴ The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu, 210046, PR China; Changzhou High-Tech Research Institute of Nanjing University and Jiangsu Target Pharma Laboratories,Inc., Changzhou, Jiangsu, 213164, PR China.

PMID: 32739350
DOI: 10.1016/j.ab.2020.113875

Abstract

mWasabi is a bright monomeric green fluorescent protein. It can be used as a fusion tag to monitor various biological events, e.g. protein localization. Here we report the selection of camelid-derived single-domain antibody fragments (nanobodies) against mWasabi. In this work, phage-display approach was employed to select the high affinity mWasabi-specific Nb (nanobodies). These nanobodies were able to recognize mWasabi or in a fused fashion with PD1. The interesting binding characteristics of these two mWasabi-specific nanobodies could be valuable for design new tools for cellular tracing or targeting based on the mWasabi-fusing protein in many different biological research fields.

Keywords: Camel; Nanobody; VHH; mWasabi fluorescent protein.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Animals
Camelidae / blood
Camelidae / immunology
Cell Surface Display Techniques / methods*
HEK293 Cells
Humans
Immunoglobulin G / blood
Luminescent Proteins / chemistry*
Luminescent Proteins / immunology
Luminescent Proteins / isolation & purification
Programmed Cell Death 1 Receptor / analysis
Programmed Cell Death 1 Receptor / metabolism
Protein Binding
Recombinant Fusion Proteins / analysis
Recombinant Fusion Proteins / metabolism
Recombinant Proteins / biosynthesis
Recombinant Proteins / immunology
Recombinant Proteins / isolation & purification
Recombinant Proteins / metabolism
Sequence Alignment
Single-Domain Antibodies / immunology*
Single-Domain Antibodies / isolation & purification*

Substances

Immunoglobulin G
Luminescent Proteins
Programmed Cell Death 1 Receptor
Recombinant Fusion Proteins
Recombinant Proteins
Single-Domain Antibodies