Abstract
Identification of core proteins carrying the CA19-9 (carbohydrate antigen, sialyl Lewis a) epitope from various tissues will improve the diagnosis of pancreatic cancer in terms of specificity and sensitivity. In this study, we attempted to identify sialyl Lewis a-carrier proteins specifically expressed in pancreatic cancer. Pancreatic cancer is difficult to detect in the early stages of the disease, resulting in a high level of mortality. Therefore, in order to determine the correct course of treatment, it is vital to distinguish cancer from obstruction of the bile duct or other diseases. Our strategy to identify the carrier proteins was as follows: glycoproteins carrying sialyl Lewis a antigen were enriched from pancreatic cancer cell lines using anti-sialyl Lewis a antibody and then subjected to Peptide Mass Fingerprinting analysis. Based on these studies we identified nine glycoproteins carrying the sialyl Lewis a epitope. We evaluated candidate molecules by biochemical analyses of culture supernatants and human sera. In particular, we focused on one candidate molecule carrying a sialyl Lewis a epitope, Galectin-3BP/MAC2BP; M2BP, which was analyzed in detail. These results verified that our candidate molecule is a core protein carrying the sialyl Lewis a epitope. Furthermore, we demonstrated sandwich ELISA, which showed that the glycoprotein was able to detect CA19-9 antigen in culture supernatants. Our approach facilitated the identification of the core protein carrying the sialyl Lewis a epitope. We believe our approach will enable future developments in cancer glycobiomarker identification.
Highlights
The presence of Natural Antibodies (NA) against EM regarding specificity and have gained increasing attention in proteome analysis for diagnosis, developing, monitoring and effective treatment of osteoarthritis of the knee
Serological identification of natural antibodies against endogenous mediators in healthy Individuals and patients with osteoarthritis of the knee by the novel ELISA Natural self-reactive antibodies of the IgM, IgG and IgA isotype are present in the serum of healthy individuals and kOA patients
Different classes of NA (IgM, IgG, IgA) against EM (BK, ANII, VEGF, bFGF) were detected with novel ELISA protocol in the sera of kOA patients as well as in the sera of healthy individuals (Table 3). These EM represent a group of novel self antigens which are targeted by NA from kOA patients
Summary
The presence of NA against EM regarding specificity and have gained increasing attention in proteome analysis for diagnosis, developing, monitoring and effective treatment of osteoarthritis of the knee (kOA). An understanding of the various regulatory systems controlling blood vessel growth, inflammation and pain in the join should lead to help explain kOA disease progression. To investigate the specific presence of NA (IgM, IgG, IgA) against EM (BK, AII, VEGF, bFGF) in the sera of kOA patients and control and to correlate this with process of joint destruction. Osteoarthritis of the knee (kOA) can be a progressive disabling disease, which results from the pathological imbalance of degradative and reparative processes, with concomitant inflammatory changes [2]. Complicated path biological interactions between the KallikreinKinin System (KKS), the Rennin-Angiotensin (RAS), angiogenesis and natural immunity could contribute to joint destruction in the disease process of kOA [15,16,17,18,19]. It is likely that biomarkers will be used in conjunction with imaging in order to establish stage of disease, predict progression, and assess improvement in the setting of clinical trials [20,21,22]
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