Synthesis and Single-cell Analysis Application of Antibody-Element-Tag Based on Peptide Affinity-mediated Site-specific Labeling Strategy
10.19756/j.issn.0253-3820.241247
- VernacularTitle:于多肽亲和介导定点偶联策略的抗体元素标签的合成及单细胞分析应用
- Author:
Lei WANG
1
;
Li YI
;
Hua-Min LI
;
Shi CHEN
;
Li-Min YANG
;
Xiao-Wen YAN
;
Qiu-Quan WANG
Author Information
1. 厦门大学化学化工学院,化学系,谱学分析与仪器教育部重点实验室,厦门 361005
- Keywords:
Peptide mediated affinity labeling;
Site-selective conjugation;
Antibodies;
Element tags;
Single cell inductively coupled plasma mass spectrometry
- From:
Chinese Journal of Analytical Chemistry
2024;52(10):1496-1507
- CountryChina
- Language:Chinese
-
Abstract:
Recently,inductively coupled plasma mass spectrometry(ICP-MS)has shown great potential in the quantitative analysis of biomolecules due to its high sensitivity,high accuracy,anti-interference ability and multi-element detection capability.Elemental tags enable specific labeling of target biomolecules with element-reporter groups,playing a critical role in the performance of bioanalysis using ICP-MS.Antibodies are highly specific to antigens on cell surfaces and are one of the most commonly used targeted delivery vehicles.Traditional random chemical coupling methods are difficult to control the coupling sites and loading numbers of element reporters,which lead to heterogeneity of antibody conjugates,affecting the specific recognition and binding ability of antibodies and antigens,and thus reducing the targeting ability and quantitative accuracy.To solve these issues,site-specific coupling of antibodies with element reporter was studied in this work.By using peptide-mediated affinity labeling technology,a short chain DNA was specifically labeled to immunoglobulin G(IgG)antibody K248,and then DNA base complementary pairing was used to realize accurate assembly of monoclonal antibodies and Eu-modified phage MS2 capsid,resulting in the preparation of highly sensitive and selective element tags.This method overcame the heterogeneity problem of element tags caused by random coupling,and ensured the targeting specificity,affinity and signal amplification ability of element tags.The element tag prepared here showed a specificity to bind/label with epidermal growth factor receptor(EGFR)biomarker on the surface of cancer cells,realizing single cancer cell analysis using ICP-MS.
