3D Visualization of Human Blood Vascular Networks Using Single-Domain Antibodies Directed against Endothelial Cell-Selective Adhesion Molecule (ESAM)

被引：2

作者：

Hansmeier, Nils Rouven ^{[1
,2
,3
]}

Bueschlen, Ina Sophie ^{[1
,2
]}

Behncke, Rose Yinghan ^{[1
,2
]}

Ulferts, Sascha ^{[1
,2
]}

Bisoendial, Radjesh ^{[4
,5
]}

Haegerling, Rene ^{[1
,2
,3
,6
]}

机构：

[1] Charite Univ Med Berlin, Inst Med & Human Genet, Res Grp Lymphovasc Med & Translat 3D Histopathol, Augustenburger Pl 1, D-13353 Berlin, Germany

[2] Charite Univ Med Berlin, BIH Ctr Regenerat Therapies, Berlin Inst Hlth, Augustenburger Pl 1, D-13353 Berlin, Germany

[3] Max Planck Inst Mol Genet, Res Grp Dev & Dis, Ihnestr 63-73, D-14195 Berlin, Germany

[4] Maasstad Hosp, Dept Rheumatol & Clin Immunol, Maasstadweg 21, NL-3079 DZ Rotterdam, Netherlands

[5] Erasmus MC, Dept Immunol, Doctor Molewaterpl 40, NL-3015 GD Rotterdam, Netherlands

[6] Charite Univ Med Berlin, Berlin Inst Hlth, BIH Acad, Clinician Scientist Program, Charitepl 1, D-10117 Berlin, Germany

来源：

INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES | 2022年 / 23卷 / 08期

关键词：

ESAM; nanobodies; single-domain antibodies; light sheet imaging; 3D microscopy; blood vessel marker; histopathology; 3D reconstruction; NANOBODIES; PROTEIN; SEQUENCE; TOOLS;

D O I：

10.3390/ijms23084369

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

High-quality three-dimensional (3D) microscopy allows detailed, unrestricted and non-destructive imaging of entire volumetric tissue specimens and can therefore increase the diagnostic accuracy of histopathological tissue analysis. However, commonly used IgG antibodies are oftentimes not applicable to 3D imaging, due to their relatively large size and consequently inadequate tissue penetration and penetration speed. The lack of suitable reagents for 3D histopathology can be overcome by an emerging class of single-domain antibodies, referred to as nanobodies (Nbs), which can facilitate rapid and superior 2D and 3D histological stainings. Here, we report the generation and experimental validation of Nbs directed against the human endothelial cell-selective adhesion molecule (hESAM), which enables spatial visualization of blood vascular networks in whole-mount 3D imaging. After analysis of Nb binding properties and quality, selected Nb clones were validated in 2D and 3D imaging approaches, demonstrating comparable staining qualities to commercially available hESAM antibodies in 2D, as well as rapid and complete staining of entire specimens in 3D. We propose that the presented hESAM-Nbs can serve as novel blood vessel markers in academic research and can potentially improve 3D histopathological diagnostics of entire human tissue specimens, leading to improved treatment and superior patient outcomes.

引用

页数：13