A novel synthetic bipartite carrier protein for developing glycotope-based vaccines

Abstract

Development of successful vaccines against glycotopes remains a major challenge. In the current studies, we have successfully developed a novel carrier protein for glycotopes based on the concept of antigen clustering and specific stimulation of T helper cells to mount strong antibody response to glycotopes. The bipartite carrier protein consists of a tandem repeat of a cysteine-rich peptide for docking of clustered glycotopes to effectively activate B cells and an Fc domain for antigen delivery to antigen presenting cells (APCs). To demonstrate its utility, we conjugated the tumor-specific monosaccharide antigen Tn to this novel carrier protein and successfully developed a Tn vaccine against cancer in animal models. The Tn vaccine effectively elicited high-titer IgG1 antibodies against Tn in immunized mice, and effectively suppressed the development of prostate cancer in Transgenic Adenocarcinoma of the Mouse Prostate (TRAMP) mice. Our results suggest that this novel bipartite carrier protein could be effectively used for developing anti-glycotope vaccines such as the anticancer Tn vaccine.

Reciprocal relationship of Tn/NF‐κB and sTn as an indicator of the prognosis of oral squamous cell carcinoma

“…Recently, we have successfully produced anti‐Tn and anti‐sTn monoclonal antibodies (mAb) with high affinity and high specificity. These two antibodies recognize their cognate antigens exclusively . In this study, we have reported the reciprocal relation of Tn and sTn in OSCC progression.…”

“…They were then treated with 0.3% H 2 O 2 for 30 min to block endogenous peroxidase; and incubated with 5% normal goat serum in phosphate‐buffered saline (PBS) for 2 h at room temperature to block non‐specific antibody reactions. After three washes with PBS containing 0.1% Tween 20 (PBST), sections were incubated with anti‐Tn (1 in 200 dilution), anti‐sTn (1 in 200 dilution) or anti‐NF‐κB antibody (1 in 200 dilution; Cell Signalling Technology, Denver, CO, USA) for 30 min at room temperature, while negative controls were incubated with PBS under the same conditions. After rinsing in PBST, tissue sections were incubated with Envision horseradish peroxidase (Dako, Glostrup, Denmark) for 30 min at room temperature and subsequently incubated with a substrate–DAB chromogen buffer (Dako), before being counterstained with Mayer's haematoxylin, dehydrated and coverslipped with mounting medium.…”

“…Anti‐Tn and ‐sTn antibodies were generated as described elsewhere . In brief, we conjugated Tn and sTn individually to a bipartite antigen carrier protein (Mw = 34 kDa).…”

“…Using anti‐Tn vaccine, we were able to obtain anti‐Tn antibodies with high specificity and high affinity. Applying anti‐Tn antibody, we have demonstrated that the expression of Tn is correlated positively with the degree of malignancy in prostate cancer . However, it is not known if Tn, sTn and NF‐κB are involved in the biological behaviour of invasive OSCC.…”

A Vaccine Targeted at CETP Alleviates High Fat and High Cholesterol Diet-Induced Atherosclerosis and Non-Alcoholic Steatohepatitis in Rabbit

“…The procedures used have been described previously [25] . In brief, levels of NF-kB and RAM-1 expression in the aorta and liver tissue were determined by immunohistochemical staining using mouse anti-NF-kB and anti-RAM-11 antibodies (Dako).…”