Generation of a human antibody that inhibits TSPAN8-mediated invasion of metastatic colorectal cancer cells

Kim, Taek Keun; Park, Chang Sik; Jeoung, Mee Hyun; Lee, Woo Ran; Go, Nam Kyung; Choi, Jong Rip; Lee, Yong Jin; Shim, Hyunbo; Lee, Sukmook

doi:10.1016/j.bbrc.2015.11.031

Cited by 22 publications

(24 citation statements)

References 16 publications

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“…Immunoblot analysis was performed as described previously with minor modifications [24]. Briefly, cell lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membranes using a wet transfer system (GE Healthcare Life Sciences, Piscataway, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

A Human Antibody That Binds to the Sixth Ig-Like Domain of VCAM-1 Blocks Lung Cancer Cell Migration In Vitro

Kim

Jang

Park

et al. 2017

IJMS

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Vascular cell adhesion molecule-1 (VCAM-1) is closely associated with tumor progression and metastasis. However, the relevance and role of VCAM-1 in lung cancer have not been clearly elucidated. In this study, we found that VCAM-1 was highly overexpressed in lung cancer tissue compared with that of normal lung tissue, and high VCAM-1 expression correlated with poor survival in lung cancer patients. VCAM-1 knockdown reduced migration of A549 human lung cancer cells into Matrigel, and competitive blocking experiments targeting the Ig-like domain 6 of VCAM-1 (VCAM-1-D6) demonstrated that the VCAM-1-D6 domain was critical for VCAM-1 mediated A549 cell migration into Matrigel. Next, we developed a human monoclonal antibody specific to human and mouse VCAM-1-D6 (VCAM-1-D6 huMab), which was isolated from a human synthetic antibody library using phage display technology. Finally, we showed that VCAM-1-D6 huMab had a nanomolar affinity for VCAM-1-D6 and that it potently suppressed the migration of A549 and NCI-H1299 lung cancer cell lines into Matrigel. Taken together, these results suggest that VCAM-1-D6 is a key domain for regulating VCAM-1-mediated lung cancer invasion and that our newly developed VCAM-1-D6 huMab will be a useful tool for inhibiting VCAM-1-expressing lung cancer cell invasion.

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Section: Methodsmentioning

confidence: 99%

A Human Antibody That Binds to the Sixth Ig-Like Domain of VCAM-1 Blocks Lung Cancer Cell Migration In Vitro

Kim

Jang

Park

et al. 2017

IJMS

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“…A biolayer interferometry (BLI) assay was performed using an Octet® RED96 system (ForteBio/Pall Life Sciences, Menlo Park, CA, USA) as described previously with minor modifications (Kim et al, 2015). CD44v3-10-Fc was immobilized onto amine-reactive biosensors (ForteBio) by amine coupling according to the manufacturer's instructions.…”

Section: Real-time Measurement Of Antibody-antigen Interactionsmentioning

confidence: 99%

Development of a sandwich enzyme-linked immunosorbent assay for the detection of CD44v3 using exon v3- and v6-specific monoclonal antibody pairs

Jeoung

Kim

Shim

et al. 2016

Journal of Immunological Methods

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“…Cell invasion assays were performed as described previously with minor modifications [20]. Briefly, 1.5 × 10 5 A549 or NCI-H1299 cells were plated in 6-well plates, allowed to attach overnight, and treated with 20 ng/ml hTNFα (R&D systems, Minneapolis, MN, USA) for 24 h at 37°C.…”

Section: Transwell Invasion Assaysmentioning

confidence: 99%

“…Immunoblot analysis was performed as described previously with minor modifications [20]. Briefly, cell lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto nitrocellulose membranes using a wet transfer system (GE Healthcare Life Sciences, Piscataway, NJ, USA).…”

Section: Immunoblot Analysismentioning

confidence: 99%

“…A biolayer interferometry (BLI) assay was performed using an Octet® RED96 system (ForteBio/Pall Life Sciences, Menlo Park, CA, USA) as described previously with minor modifications [20]. Briefly, hVCAM-1-ext or mVCAM-1-ext was immobilized onto amine-reactive biosensors (ForteBio) by amine coupling according to the manufacturer's instructions.…”

Section: Real-time Measurement Of Antibody-antigen Interactionsmentioning

confidence: 99%

See 1 more Smart Citation

Identification of the Sixth Ig-like Domain of VCAM-1 as a Novel Therapeutic Target in Lung Cancer Cell Invasion

Kim

Jang

Park

et al. 2017

Preprint

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Vascular cell adhesion molecule-1 (VCAM-1) is closely associated with tumor progression and metastasis. However, the relevance and role of VCAM-1 in lung cancer have not been clearly elucidated. In this study, we found that VCAM-1 was highly overexpressed in lung cancer tissue compared with that of normal lung, and high VCAM-1 expression correlated with poor survival of lung cancer patients. VCAM-1 knockdown reduced invasion in A549 human lung cancer cells, and competitive blocking experiments targeting the Ig-like domain 6 of VCAM-1 (VCAM-1-D6) demonstrated that the VCAM-1-D6 domain was critical for VCAM-1-mediated A549 cell invasion. Next, we developed a human monoclonal antibody specific to human and mouse VCAM-1-D6 (VCAM-1-D6 huMab), which was isolated from a human synthetic antibody library using phage display technology. Finally, we showed that VCAM-1-D6 huMab had a nanomolar affinity for VCAM-1-D6 and that it potently suppressed invasion in A549 and NCI-H1299 lung cancer cell lines. Taken together, these results suggest that VCAM-1-D6 is a novel therapeutic target in VCAM-1-mediated lung cancer invasion and that our newly developed VCAM-1-D6 huMab will be a useful tool for inhibiting VCAM-1-expressing lung cancer cell invasion.

show abstract

Generation of a human antibody that inhibits TSPAN8-mediated invasion of metastatic colorectal cancer cells

Cited by 22 publications

References 16 publications

A Human Antibody That Binds to the Sixth Ig-Like Domain of VCAM-1 Blocks Lung Cancer Cell Migration In Vitro

A Human Antibody That Binds to the Sixth Ig-Like Domain of VCAM-1 Blocks Lung Cancer Cell Migration In Vitro

Development of a sandwich enzyme-linked immunosorbent assay for the detection of CD44v3 using exon v3- and v6-specific monoclonal antibody pairs

Identification of the Sixth Ig-like Domain of VCAM-1 as a Novel Therapeutic Target in Lung Cancer Cell Invasion

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