A High-Affinity Repebody for Molecular Imaging of EGFR-Expressing Malignant Tumors

Yun, Miyong; Kim, Dong Yeon; Lee, Joong-Jae; Kim, Hyeon-Sik; Kim, Hyung-Seok; Pyo, Ayoung; Ryu, Yiseul; Kim, Tae Yoon; Zheng, Jin; Yoo, Su Woong; Hyun, Hoon; Oh, Gyungseok; Jeong, Ji Won; Moon, Myeong-Ju; Min, Jo Young; Kwon, Seong; Kim, Jung Young; Chung, Euiheon; Hong, Yeongjin; Lee, Wan‐Sik; Kim, Kwang Ho; Min, Jung‐Joon

doi:10.7150/thno.18096

Cited by 25 publications

(14 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Lampreys last shared a common ancestor with mammals~550 million years ago, and this evolutionary distance may provide the potential for the generation of VLRB proteins that recognize highly conserved proteins and carbohydrates that are not immunogenic in rodents 18 . These reagents have proven useful in a number of critical biological assay platforms, such as glycan microarray, flow cytometry, western blotting, and in probing tissue in immunohistochemistry assays 51,52 . VLRB proteins also have a wide variety of applications in biomedical research, including inhibition of cell signaling pathways 53,54 , as scaffolds for drug delivery 55 , and as diagnostic reagents 11,21,23 .…”

Section: Discussionmentioning

confidence: 99%

Development of smart anti-glycan reagents using immunized lampreys

et al. 2020

View full text Add to dashboard Cite

Studies on the expression of cellular glycans are limited by a lack of sensitive tools that can discriminate specific structural features. Here we describe the development of a robust platform using immunized lampreys (Petromyzon marinus), which secrete variable lymphocyte receptors called VLRBs as antibodies, for generating libraries of anti-glycan reagents. We identified a wide variety of glycan-specific VLRBs detectable in lamprey plasma after immunization with whole fixed cells, tissue homogenates, and human milk. The cDNAs from lamprey lymphocytes were cloned into yeast surface display (YSD) libraries for enrichment by multiple methods. We generated VLRB-Ig chimeras, termed smart anti-glycan reagents (SAGRs), whose specificities were defined by microarray analysis and immunohistochemistry. 15 VLRB antibodies were discovered that discriminated between linkages, functional groups and unique presentations of the terminal glycan motif. The development of SAGRs will enhance future studies on glycan expression by providing sequenced, defined antibodies for a variety of research applications.

show abstract

Section: Discussionmentioning

confidence: 99%

Development of smart anti-glycan reagents using immunized lampreys

et al. 2020

View full text Add to dashboard Cite

show abstract

“…To increase the stability and recombinant expression yield of the VLR scaffold, two groups have created consensus VLR scaffolds, dVLRs, and repebodies, which can be expressed in E coli and have been used to target proteins involved in a variety of diseases . These scaffolds are highly stable, and have shown general applicability to detect a target of interest by high‐throughput screening …”

Section: Engineered Binding Scaffoldsmentioning

confidence: 99%

Engineering repeat proteins of the immune system

McCord

Grove

2020

Biopolymers

View full text Add to dashboard Cite

Limitations associated with immunoglobulins have motivated the search for novel binding scaffolds. Repeat proteins have emerged as one promising class of scaffolds, but often are limited to binding protein and peptide targets. An exception is the repeat proteins of the immune system, which have in recent years served as an inspiration for binding scaffolds which can bind glycans and other classes of biomolecule.Like other repeat proteins, these proteins can be very stable and have a monomeric mode of binding, with elongated and highly variable binding surfaces. The ability to target glycans and glycoproteins fill an important gap in current tools for research and biomedical applications.

show abstract

“…The stability of 18 F-DMFB was analyzed by modification of a previously reported method (28). Instant thin-layer chromatography silica-gel strips were developed with dichloromethane and methanol (10:1) and were scanned.…”

Section: Stability Study and In Vitro Binding Assaymentioning

confidence: 99%

N-(2-(Dimethylamino)Ethyl)-4-¹⁸F-Fluorobenzamide: A Novel Molecular Probe for High-Contrast PET Imaging of Malignant Melanoma

Pyo

Kim

et al. 2018

J Nucl Med

Self Cite

View full text Add to dashboard Cite

Malignant melanoma is an aggressive and serious form of skin cancer, with prognosis and treatment outcome depending heavily on the clinical stage of the disease at the time of diagnosis. Here, we synthesized a novel 18 F-labeled benzamide derivative to target melanoma and then evaluated its biologic characteristics in small-animal models. Methods: N-(2-(dimethylamino)ethyl)-4-18 F-fluorobenzamide ( 18 F-DMFB) was synthesized by reaction of N-succinimidyl 4-18 Ffluorobenzoate with N,N-dimethylethylenediamine. The binding affinity of 18 F-DMFB was measured in B16F10 (mouse melanoma) cells with or without L-tyrosine. Small-animal PET imaging with 18 F-DMFB was performed on B16F10 xenograft and metastasis mouse models. Results: The overall non-decay-corrected radiochemical yield of 18 F-DMFB was approximately 10%-15%. Uptake of 18 F-DMFB was melanin-specific, as cellular uptake in B16F10 increased more than 18-fold in the presence of L-tyrosine. Biodistribution studies revealed that 18 F-DMFB accumulated, and was retained, in B16F10 xenografts for 120 min (10, 30, 60, and 120 min: 9.24, 10.80, 13.0, and 10.59 percentage injected dose/g, respectively) after radiotracer injection. Liver uptake of 18 F-DMFB decreased from 10 to 120 min and showed fast clearance (10, 30, 60, and 120 min: 11.19, 5.7, 2.47, and 0.4 percentage injected dose/g). Furthermore, 18 F-DMFB allowed visualization of metastatic lesions immediately after injection and was retained in lesions for over 60 min, with a high tumor-to-background ratio. Conclusion: 18 F-DMFB demonstrated a high melanin-targeting ability and tumor-specific tumor uptake in both primary and metastatic lesions in animal models bearing malignant melanoma. 18 F-DMFB may be a potential PET imaging agent for melanoma.

show abstract

A High-Affinity Repebody for Molecular Imaging of EGFR-Expressing Malignant Tumors

Cited by 25 publications

References 40 publications

Development of smart anti-glycan reagents using immunized lampreys

Development of smart anti-glycan reagents using immunized lampreys

Engineering repeat proteins of the immune system

N-(2-(Dimethylamino)Ethyl)-4-¹⁸F-Fluorobenzamide: A Novel Molecular Probe for High-Contrast PET Imaging of Malignant Melanoma

Contact Info

Product

Resources

About

A High-Affinity Repebody for Molecular Imaging of EGFR-Expressing Malignant Tumors

Cited by 25 publications

References 40 publications

Development of smart anti-glycan reagents using immunized lampreys

Development of smart anti-glycan reagents using immunized lampreys

Engineering repeat proteins of the immune system

N-(2-(Dimethylamino)Ethyl)-4-18F-Fluorobenzamide: A Novel Molecular Probe for High-Contrast PET Imaging of Malignant Melanoma

Contact Info

Product

Resources

About

N-(2-(Dimethylamino)Ethyl)-4-¹⁸F-Fluorobenzamide: A Novel Molecular Probe for High-Contrast PET Imaging of Malignant Melanoma