Novel monoclonal antibodies against pancreatic juice from pancreatic cancer patients and their possible application in differential diagnosis

Abstract

Abstract. Pancreatic cancer (PC) has a poor clinical prognosis with a <10% 5-year survival rate. Because there are no specific biomarkers of PC, it is difficult to detect small PC tumors and most patients are diagnosed at an advanced stage. Specific biomarkers are useful tools for the early detection of cancer. However, PC-related biomarkers, such as CA19-9 lack specificity and sensitivity. In this study, we took an immunological approach to establish novel monoclonal antibodies (mAbs) specific for the pancreatic juice from PC patients, which would be potentially useful in the diagnosis of PC. Mice were immunized by subtractive immunization using mixed pancreatic juices from chronic pancreatitis and PC patients as the tolerogen and the immunogen, respectively. After screening by Western blotting, four mAbs were obtained: 2P-1-2-1, 2P-1-17-1, 6P-3-2-4 and 7P-9-11-6. The mAb 2P-1-2-1 showed reactivity against the tolerogen at 115 and 120 kDa, but only the 120-kDa antigen was also reactive to the immunogen. The mAb 2P-1-17-1 showed an intense smear reactivity at ~150 kDa against the immunogen. Finally, the mAbs 6P-3-2-4 and 7P-9-11-6 showed PC-specific reactivity to the immunogen at >250 kDa and at ~70 kDa, respectively. We propose that investigation of pancreatic juice samples with these mAbs may enable us to perform reliable differential diagnosis of benign and malignant diseases. Furthermore, we demonstrated that subtractive immunization is a useful method for producing mAbs specific for the pancreatic juice from PC patients.

Enhanced immunization techniques to obtain highly specific monoclonal antibodies

“…Almost 20 y after this technique was described, Hamabashiri et al . 8 produced 4 mAbs (2P-1–2–1, 2P-1–17–1, 6P-3–2–4 and 7P-9–11–6) specific to pancreatic juice from cancer patients, which have a potential application for differential diagnosis. Their immunization protocol was performed using female BALB/c mice, 5–6 weeks old, tolerized with 25 µg of inflammatory pancreatic juice diluted in phosphate-buffered saline (PBS) and adjuvant.…”

“…Hamabashiri et al. 8 showed that their approach is an effective technique to obtain specific mAbs to rare or poor immunogenic antigens, and that it can be used to provide tools for new strategies in the premature diagnosis of cancer.…”