The α-enolase, a major form of enolase, was found in the early stages of embryonic development with multiple roles involving in enzymatic, structural, and receptor functions. In addition to its roles in autoimmune disorders and bacterial infection, α-enolase expression was up-regulated in several highly tumorigenic or metastatic cell lines, implying it may play a role in tumor adhesion and invasion. Moreover, α-enolase expression showed significant correlation with tumorigenicity in several types of cancer, which suggests its pathophysiologic role in cancer formation. It was previously reported that up-regulation of α-enolase protein was detected in 65% of patients with non-small cell lung cancers (NSCLC). In contrast, a high titer of anti-α-enolase antibodies was developed in a smaller proportion (7.4%) of these patients than in non-tumor-associated patients and healthy subjects. The results suggested that antibodies that can specifically recognize α-enolase protein would be very useful not only as tools to study how α-enolase expression may be involved in the tumor formation, but also as potential diagnostic and therapeutic agents for cancer cell proliferation and metastasis in clinical application. Recently, nano-technology has been widely applied in many research fields. Accordingly, we will generate nano-particles with surface-modified antibody probe to detect the presence of α-enolase protein in the patient sera. So far, we have generated and characterized polyclonal IgY antibodies (Fig. 1 and 2) and 2 scFv antibodies (Fig. 3 to 8), which however need to be further confirmed. We are also in the process of production of monoclonal anti-α-enolase antibodies (Fig. 9). Thus, in the present study, we propose 1. to clone and express the human α-enolase protein gene in E. coli 2. to generate and characterize polyclonal and single-chain variable fragment (scFv) anti-α-enolase antibodies from immunized mice and chickens 3. to investigate the potential inhibitory effect of these antibodies on tumorigenesis 4. to study the mechanisms involved in the inhibitory function of these antibodies 5. to develop a highly sensitive method for detecting the presence of α-enolase protein or anti-α-enolase antibodies in the patient sera 6. to humanize the anti-α-enolase antibodies with high affinity through antibody engineering technology
|Effective start/end date
|8/1/12 → 7/31/13
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