Application of Single Domain Antibody in Biomedical Imaging (Invited)

Zhou, Siyu; Luo, Yunhe; Zeng, Yiqi; Yang, Yicheng; Yu, Yongbo; Wu, Changfeng

2024-08-01

10.3788/CJL240698

CHINESE JOURNAL OF LASERS-ZHONGGUO JIGUANG   影响因子和分区

0258-7025

["Significance Antibodies have potential for use in clinical diagnostics, therapeutics, and biomedical research. Since the approval of the first monoclonal antibody- associated drug by the United States Food and Drug Administration (FDA) in 1986, monoclonal antibodies have been considered the most promising targeted agents against diseases. Over the past four decades, an increasing number of monoclonal antibodies have been used in clinical settings and preclinical trials. Examples include daratumumab, which targets CD38 in multiple myeloma cells, and nivolumab, which reactivates T- cell recognition ability by inhibiting the PD-1/PD-L1- 1/PD- L1 pathway and preventing cancer progression. However, large molecular monoclonal antibodies (molecular mass of--150 kDa) have complex structures and physicochemical properties, which lead to in vivo limitations such as low tissue penetration, extended circulation, and slow clearance. In addition, high- price production and potential immunogenicity issues imped the development and application of monoclonal antibody- based imaging and therapeutic agents. A single domain antibody is a small antibody fragment first discovered in Camelidae, , which can effectively recognize antigens of interest. Compared with conventional monoclonal antibodies, single domain antibodies exhibit superior properties, such as small size (--15 kDa), high affinity, high stability, low immunogenicity, and ease of tissue penetration, making them promising candidates for targeted imaging and drug delivery in research and clinical applications. This review systematically introduces progress in the research and application of single domain antibody- based probes in live- cell imaging, super- resolution imaging, in vivo fluorescence, and nuclear imaging. This review also discusses the future opportunities and challenges for single- domain antibodies in biomedicine. Progress The applications of single domain antibodies in cell imaging, including live- cell imaging and super- resolution imaging, were first introduced. Rothbauer et al. first investigated the construction of a chromobody in 2006 by fusing the gene sequences of a single domain antibody and a fluorescent protein, enabling its expression in live cells and binding to antigens of interest (Fig. 2). Subsequently, various chromobodies that target finer subcellular structures, such as organelle membrane actin, nuclear actin, and vimentin, have been developed. To improve the controllability and applicability of single domain antibodies in complex cellular environments, O'shea et al. developed a photocaged probe (Fig. 3). For super- resolution imaging, single- domain antibodies can be used to label subcellular structures with high densities and minimal linkage errors. The single domain antibody probes and labeling strategies for single molecule localization microscopy (SMLM), stochastic optical reconstruction microscopy (STORM), stimulated emission depletion microscopy (STED), and expansion microscopy (ExM) are summarized in the second section (Figs. 4 and 5). Jungmann et al. introduced a DNA barcoding method called resolution enhancement by sequential imaging (RESI), which improves the resolution of fluorescence microscopy to the & Aring;ngstr & ouml;m scale using single domain antibodies (Fig. 6). In the third section, the bioimaging applications of single domain antibody probes are presented to demonstrate their advantages, including short circulation time, high affinity, deep tissue penetration, and rapid enrichment.","Single domain antibodies conjugated with near- infrared fluorescent dyes have become effective for tumor imaging and image- guided surgery (Fig. 7). When combined with short half-life- life nuclides, such as 68 Ga, 18 F, and 64 Cu, single domain antibody- based tracers demonstrate high specificity and low radiation risk in immunoPET and immunoSPECT imaging, which are better than the traditional 18 F- FDG (Fig. 8). Conclusions and Prospects The advantages of single domain antibodies, including their small size, high stability, deep penetration, renal clearance, and low immunogenicity, are promising for biomedical applications. In cell imaging, single domain antibodies can be effectively modified using various tags, dyes, and adapters, making them versatile tools for live- cell and super- resolution imaging. At the organismal level, single domain antibodies exhibit a short circulation time, rapid clearance, and low risk of toxicity and immunogenicity, which facilitate real-time- time and highly specific in vivo imaging. Current preclinical data indicate that single domain antibody- associated probes have a tremendous translational potential. The first radiolabeled single domain antibody probe, 2Rs15d (HER2-targeting),- targeting), labeled with either 68 Ga or 131 I, has entered clinical phase I trials. Numerous studies have been conducted on the design and screening of single domain antibodies that target various antigens. However, it is crucial to develop novel and effective biomarkers and screen for the corresponding high- specific single domain antibodies for the diagnosis and treatment of complex diseases, particularly heterogeneous tumors. The key to designing single domain antibody probes is exploring stable, universal, and controllable strategies for modification using dyes, chelators, and other labels. Because of the small size and limited active sites of single domain antibodies, the effects of conjugation sites, labeling methods, and properties of tags on single domain antibodies are significantly more pronounced than those on monoclonal antibodies. Although random labeling methods that rely on endogenous cysteine or lysine residues in proteins are simple and widely used, they may form heterogeneous products with variable functionalization ratios, and sometimes lead to a loss of targeting ability. Enzymes, including sortase-and- and microbial transglutaminase-mediated- mediated strategies and click chemistry, may be effective and promising approaches for constructing single domain antibody- based probes and their clinical translation. However, challenges remain regarding their application and improvement. In addition, advancements in chelators, isotopes, fluorescent dyes, imaging techniques, and other fields will continue to promote the development and translation of single domain antibody probes."]

medical optics single domain antibody biomedical imaging fluorescence imaging

ESCI ; EI

英语

第一 ; 通讯

Optics

Optics

WOS:001301555100009

CHINESE LASER PRESS

Web of Science

Southern Univ Sci & Technol, Dept Biomed Engn, Guangdong Prov Key Lab Adv Biomat, Shenzhen 518055, Guangdong, Peoples R China

Zhou, Siyu,Luo, Yunhe,Zeng, Yiqi,et al. Application of Single Domain Antibody in Biomedical Imaging (Invited)[J]. CHINESE JOURNAL OF LASERS-ZHONGGUO JIGUANG,2024,51(15).

Zhou, Siyu,Luo, Yunhe,Zeng, Yiqi,Yang, Yicheng,Yu, Yongbo,&Wu, Changfeng.(2024).Application of Single Domain Antibody in Biomedical Imaging (Invited).CHINESE JOURNAL OF LASERS-ZHONGGUO JIGUANG,51(15).

Zhou, Siyu,et al."Application of Single Domain Antibody in Biomedical Imaging (Invited)".CHINESE JOURNAL OF LASERS-ZHONGGUO JIGUANG 51.15(2024).