Development of Human Monoclonal Antibody for Claudin-3 Overexpressing Carcinoma Targeting

Abstract

Most malignant tumors originate from epithelial tissues in which tight junctions mediate cell–cell interactions. Tight junction proteins, especially claudin-3 (CLDN3), are overexpressed in various cancers. Claudin-3 is exposed externally during tumorigenesis making it a potential biomarker and therapeutic target. However, the development of antibodies against specific CLDN proteins is difficult, because CLDNs are four-transmembrane domain proteins with high homology among CLDN family members and species. Here, we developed a human IgG1 monoclonal antibody (h4G3) against CLDN3 through scFv phage display using CLDN3-overexpressing stable cells and CLDN3-embedded lipoparticles as antigens. The h4G3 recognized the native conformation of human and mouse CLDN3 without cross-reactivity to other CLDNs. The binding kinetics of h4G3 demonstrated a sub-nanomolar affinity for CLDN3 expressed on the cell surface. The h4G3 showed antibody-dependent cellular cytotoxicity (ADCC) according to CLDN3 expression levels in various cancer cells by the activation of FcγRIIIa (CD16a). The biodistribution of h4G3 was analyzed by intravenous injection of fluorescence-conjugated h4G3 which showed that it localized to the tumor site in xenograft mice bearing CLDN3-expressing tumors. These results indicate that h4G3 recognizes CLDN3 specifically, suggesting its value for cancer diagnosis, antibody-drug conjugates, and potentially as a chimeric antigen receptor (CAR) for CLDN3-expressing pan-carcinoma.

Antibody-Based Targeting of Interferon-Beta-1a Mutein in HER2-Positive Cancer Enhances Antitumor Effects Through Immune Responses and Direct Cell Killing

“…Because activating NK cells rapidly downregulate the expression of Fc gamma receptor III-A (CD16a) ( Peruzzi et al, 2013 ; Romee et al, 2013 ), co-culture with NK cells from PBMCs did not show sufficient ADCC effect by trastuzumab. To validate the ADCC effect by NK cells, we measured ADCC activity using NK-92MI-CD16 cell line, which is a genetically engineered cell line with sufficient activity ( Yang et al, 2020 ). Treatment of trastuzumab-R27T showed a similar ADCC effect as that of trastuzumab in HER2-positive cancer cell lines, suggesting that trastuzumab-R27T is capable of causing significant ADCC ( Figure 4C ).…”

“…Target cells were seeded in 96-well plates at a density of 2 × 10 4 cells/well and incubated overnight. The NK-92MI-CD16a cells, which stably express CD16a, were previously established and used as effector cells ( Yang et al, 2020 ). The target cells were incubated with control IgG, IFN-β-R27T, or trastuzumab-R27T at a final concentration of up to 10,000 nM and 8 × 10 4 effector cells in a CO 2 incubator for 4 h at 37°C (E:T ratio = 4:1).…”

The Adoption of Viral Capsid-Derived Virus-Like Particles (VLPs) for Disease Prevention and Treatments

“…Another important feature in the field of VLPs, was the adoption of lipoparticles. Lipoparticles are purified and homogeneous VLPs that are engineered to contain the intact membrane proteins and are useful for delivering target therapies [ 5 ]. Furthermore, VLPs displaying the alpha-Gal carbohydrate, a molecule eliciting protective immune response against multiple pathogens [ 6 ], have shown protection against Leishmaniosis [ 7 ].…”

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“…cell line, which is a genetically engineered cell line with sufficient activity (Yang et al, 2020). Treatment of trastuzumab-R27T showed a similar ADCC effect as that of trastuzumab in HER2positive cancer cell lines, suggesting that trastuzumab-R27T is capable of causing significant ADCC ( Figure 4C).…”