| 项目编号 |
PRJCA018693 |
| 项目标题 |
Modular DNA Barcoding of Nanobodies Enables Multiplexed in situ Protein Imaging and High-throughput Biomolecule Detection |
| 涉及领域 |
Medical |
| 数据类型 |
Barcoded reads
|
| 物种名称 |
unidentified
|
| 描述信息 |
The antibody-antigen interaction-based immunodetection methods are routinely used in biological and clinical laboratories. DNA-labeled antibodies have the advantage of detecting multiple target molecules simultaneously via orthogonal DNA barcode sequences. However, current antibody-DNA conjugation commonly uses non-site-specific modifications tailor-made for each specific application, which is laborious and often compromises antibody affinity and specificity. To overcome these limitations, here we introduce a simple, precise, and cost-efficient DNA barcoding strategy - multiplexed and modular barcoding of antibodies (MaMBA). Via nanobodies as the modular adaptor, MaMBA enables direct site-specific labeling to off-the-shelf IgG antibodies with one-component design. We first used MaMBA to develop the misHCR method for highly multiplexed in situ protein imaging via orthogonal hybridization chain reaction (HCR). A cleavable version of MaMBA allows iterative employment of the same HCR probes sets and thus further enhances the multiplexity of misHCR (misHCRn). misHCRn achieves simultaneous labeling of 12 different target molecules within the same mouse brain sections. We next extended the cleavable MaMBA to develop the barcode-linked immunosorbent assay (BLISA) for multiplexed and high-throughput biomolecule detections. By combining BLISA and the next-generation sequencing, we successfully measured SARS-CoV-2 IgG and HBV-associated antigens in a large number of human serum samples. Thus, MaMBA offers a highly modular and easily adaptable approach for antibody DNA-barcoding, which can be broadly applied in basic research and clinical diagnostics. |
| 样品范围 |
Barcoded reads |
| 发布日期 |
2025-07-17 |
| 出版信息 |
| PubMed ID |
文章标题 |
杂志名称 |
Doi |
发表年份 |
| 40694500
|
Modular DNA barcoding of nanobodies enables multiplexed in situ protein imaging and high-throughput biomolecule detection eLife Assessment Reviewer #1 (Public review): Author response
|
eLife
|
10.7554/eLife.105225
|
2025
|
|
|
| 项目资金来源 |
| 机构 |
项目类型 |
授权项目ID |
授权项目名称 |
| Ministry of Science and Technology China Brain Initiative Grant
|
|
2021ZD0202803
|
|
| Research Unit of Medical Neurobiology at Chinese Academy of Medical Sciences
|
|
2019RU003
|
|
|
|
| 提交者 |
Ruiyu
Wang (wangruiyu@cibr.ac.cn)
|
| 提交单位 |
Chinese Institute for Brain Research |
| 提交日期 |
2023-07-30 |
