The antibodies were proved that they were specific to the biomarkers. Therefore the antibodies can be used for the application in immunoblotting assay toward cell lysate of hepatoma cell, immunocytochemistry, immunohistochemistry, and even in real-sample detection. Additionally, the antibody can be tried for testing its anti-tumor activity. Also, the antibodies can be conjugated with a signal reporter such as a radioactive substance or other reporter to judge the location of tumors. These antibodies have the potential for the application in target therapy.
The low sensitivity of our system may due to low efficiency of conjugation. To improve the efficiency of the conjugation of AP to antibody, different parameters such as temperatures and concentration of glutaraldehyde may be applied in the reaction. Besides, different crosslinking reagents such as BS3 and N-succini-midyl 3-(2-pyridyldithio) propionate could be applied in conjugation.
References
Abelev, G. I. (1971). "Alpha-fetoprotein in ontogenesis and its association with malignant tumors." Adv Cancer Res 14: 295-358.
Ai-Ping, F., Q. Yue, et al. (2007). "A case report of remote cutaneous metastasis from male breast carcinoma." Int J Dermatol 46(7): 738-739.
Alon, T., I. Hemo, et al. (1995). "Vascular endothelial growth factor acts as a survival factor for newly formed retinal vessels and has implications for retinopathy of prematurity."
Nat Med 1(10): 1024-1028.
Arrieta, O., B. Cacho, et al. (2007). "The progressive elevation of alpha fetoprotein for the diagnosis of hepatocellular carcinoma in patients with liver cirrhosis." Bmc Cancer 7:
28.
Aschaffenburg, R. and J. E. C. Mullen (1949). "381. A rapid and simple phosphatase test for milk." Journal of Dairy Research 16(01): 58-67.
Boismenu, R., D. Semeniuk, et al. (1997). "Purification and Characterization of Human and Mouse Recombinant Alpha-Fetoproteins Expressed inEscherichia coli." Protein Expression and Purification 10(1): 10-26.
Butler-Ransohoff, J. E., D. A. Kendall, et al. (1988). "Use of site-directed mutagenesis to elucidate the role of arginine-166 in the catalytic mechanism of alkaline phosphatase."
Proceedings of the National Academy of Sciences 85(12): 4276-4278.
Carlini, P., P. Ferranti, et al. (2007). "Purification and characterization of Alpha-Fetoprotein from the human hepatoblastoma HepG2 cell line in serum-free medium." BioMetals
20(6): 869-878.
Chaidaroglou, A., D. J. Brezinski, et al. (1988). "Function of arginine-166 in the active site of Escherichia coli alkaline phosphatase." Biochemistry 27(22): 8338-8343.
Chen, L., D. Neidhart, et al. (1992). "3-D structure of a mutant (Asp101 → Ser) of E.coli alkaline phosphatase with higher catalytic activity." Protein Engineering 5(7):
605-610.
Cheng, H. T., Y. H. Chang, et al. (2007). "AFP-L3 in chronic liver diseases with persistent elevation of alpha-fetoprotein." J Chin Med Assoc 70(8): 310-317.
Daniele, B., A. Bencivenga, et al. (2004). "Alpha-fetoprotein and ultrasonography screening for hepatocellular carcinoma." Gastroenterology 127(5 Suppl 1): S108-112.
de Carlos, A., D. Montenegro, et al. (2003). "Purification of human alpha-l-fucosidase precursor expressed in Escherichia coli as a glutathione S-transferase fusion protein."
Journal of Chromatography B 786(1–2): 7-15.
Dealwis, C. G., L. Chen, et al. (1995). "3-D structure of the D153G mutant of Escherichia coli alkaline phosphatase: an enzyme with weaker magnesium binding and increased catalytic activity." Protein Engineering 8(9): 865-871.
Deugnier, Y., V. David, et al. (1984). "Serum alpha-L-fucosidase: a new marker for the diagnosis of primary hepatic carcinoma?" Hepatology 4(5): 889-892.
Deutsch, H. F. (1991). "Chemistry and biology of alpha-fetoprotein." Adv Cancer Res 56:
253-312.
Di Bisceglie, A. M., R. K. Sterling, et al. (2005). "Serum alpha-fetoprotein levels in patients with advanced hepatitis C: results from the HALT-C Trial." J Hepatol 43(3): 434-441.
el-Houseini, M. E., M. S. Mohammed, et al. (2005). "Enhanced detection of hepatocellular carcinoma." Cancer control : journal of the Moffitt Cancer Center 12(4): 248-253.
el-Houseini, M. E., M. S. Mohammed, et al. (2005). "Enhanced detection of hepatocellular carcinoma." Cancer Control 12(4): 248-253.
El-Serag, H. B., J. A. Marrero, et al. (2008). "Diagnosis and Treatment of Hepatocellular Carcinoma." Gastroenterology 134(6): 1752-1763.
Fukushima, H., J. Nishimoto, et al. (1990). "Sequencing and expression of a full-length cDNA for human alpha-L-fucosidase." J Inherit Metab Dis 13(5): 761-765.
Giardina, M. G., M. Matarazzo, et al. (1998). "Serum alpha-L-fucosidase activity and early detection of hepatocellular carcinoma: a prospective study of patients with cirrhosis."
Cancer 83(12): 2468-2474.
Gomaa, A. I., S. A. Khan, et al. (2009). "Diagnosis of hepatocellular carcinoma." World J Gastroenterol 15(11): 1301-1314.
Gorin, M. B., D. L. Cooper, et al. (1981). "The evolution of alpha-fetoprotein and albumin. I.
A comparison of the primary amino acid sequences of mammalian alpha-fetoprotein and albumin." J Biol Chem 256(4): 1954-1959.
Hauser, S. and H. A. Weich (1993). "A Heparin-Binding Form of Placenta Growth Factor (PlGF-2) is Expressed in Human Umbilical Vein Endothelial Cells and in Placenta."
Growth Factors 9(4): 259-268.
Holtz, K. M. and E. R. Kantrowitz (1999). "The mechanism of the alkaline phosphatase reaction: insights from NMR, crystallography and site-specific mutagenesis." FEBS Letters 462(1–2): 7-11.
Hooper, L. V. and J. I. Gordon (2001). "Glycans as legislators of host–microbial interactions:
spanning the spectrum from symbiosis to pathogenicity." Glycobiology 11(2):
1R-10R.
Janeway, C. M. L., X. Xu, et al. (1993). "Magnesium in the active site of Escherichia coli alkaline phosphatase is important for both structural stabilization and catalysis."
Biochemistry 32(6): 1601-1609.
Kam, W., E. Clauser, et al. (1985). "Cloning, sequencing, and chromosomal localization of human term placental alkaline phosphatase cDNA." Proceedings of the National Academy of Sciences 82(24): 8715-8719.
Kansas, G. (1996). "Selectins and their ligands: current concepts and controversies." Blood 88(9): 3259-3287.
Kew, M. (1974). "Alpha-fetoprotein in primary liver cancer and other diseases." Gut 15(10):
814-821.
Khunsook, S., J. A. Alhadeff, et al. (2002). "Purification and characterization of human seminal plasma α-l-fucosidase." Molecular Human Reproduction 8(3): 221-227.
Kim, E. E. and H. W. Wyckoff (1991). "Reaction mechanism of alkaline phosphatase based on crystal structures: Two-metal ion catalysis." Journal of Molecular Biology 218(2):
449-464.
Lamszus, K., U. Ulbricht, et al. (2003). "Levels of soluble vascular endothelial growth factor (VEGF) receptor 1 in astrocytic tumors and its relation to malignancy, vascularity, and VEGF-A." Clin Cancer Res 9(4): 1399-1405.
Le Du, M. H., T. Stigbrand, et al. (2001). "Crystal Structure of Alkaline Phosphatase from
Human Placenta at 1.8 Å Resolution." Journal of Biological Chemistry 276(12):
9158-9165.
Le, H. V. and P. P. Trotta (1991). "Purification of secreted recombinant proteins from Escherichia coli." Bioprocess Technol 12: 163-181.
Lowe, J. B. (1993). "The blood group-specific human glycosyltransferases." Baillieres Clin Haematol 6(2): 465-492.
Malaguarnera, G., M. Giordano, et al. (2010). "Serum markers of hepatocellular carcinoma."
Dig Dis Sci 55(10): 2744-2755.
Mandecki, W., M. A. Shallcross, et al. (1991). "Mutagenesis of conserved residues within the active site of Escherichia coli alkaline phosphatase yields enzymes with increased kcat." Protein Engineering 4(7): 801-804.
Maxam, A. M. and W. Gilbert (1980). "Sequencing end-labeled DNA with base-specific chemical cleavages." Methods Enzymol 65(1): 499-560.
Mizejewski, G. J. (2001). "Alpha-fetoprotein Structure and Function: Relevance to Isoforms, Epitopes, and Conformational Variants." Experimental Biology and Medicine 226(5):
377-408.
Morinaga, T., M. Sakai, et al. (1983). "Primary structures of human alpha-fetoprotein and its mRNA." Proceedings of the National Academy of Sciences 80(15): 4604-4608.
Mousa, N., Y. Gad, et al. (2012). "Increased alpha-Fetoprotein Predicts Steatosis among Patients with Chronic Hepatitis C Genotype 4." Int J Hepatol 2012: 636392.
Murphy, J. E., X. Xu, et al. (1993). "Conversion of a magnesium binding site into a zinc binding site by a single amino acid substitution in Escherichia coli alkaline
phosphatase." Journal of Biological Chemistry 268(29): 21497-21500.
Opheim, D. J. and O. Touster (1977). "The purification and characterization of rat liver lysosomal alpha-L-fucosidase." J Biol Chem 252(2): 739-743.
Orpana, A. and P. Salven (2002). "Angiogenic and Lymphangiogenic Molecules in Hematological Malignancies." Leukemia & Lymphoma 43(2): 219-224.
Pang, R. W., J. W. Joh, et al. (2008). "Biology of hepatocellular carcinoma." Ann Surg Oncol 15(4): 962-971.
Park, Y. N., Y. B. Kim, et al. (2000). "Increased expression of vascular endothelial growth factor and angiogenesis in the early stage of multistep hepatocarcinogenesis."
Archives of Pathology & Laboratory Medicine 124(7): 1061-1065.
Peters, E. H., S. Nishi, et al. (1979). "In Vitro Synthesis of Murine Pre-α-fetoprotein." Cancer Research 39(9): 3702-3706.
Rapaccini, G. L., M. Pompili, et al. (2004). "Hepatocellular carcinomas <2 cm in diameter complicating cirrhosis: ultrasound and clinical features in 153 consecutive patients."
Liver International 24(2): 124-130.
Reglero, A. and J. A. Cabezas (1976). "Glycosidases of molluscs. Purification and properties of alpha-L-fucosidase from Chamelea gallina L." Eur J Biochem 66(2): 379-387.
Schwartz, J. H. (1963). "THE PHOSPHORYLATION OF ALKALINE PHOSPHATASE."
Proceedings of the National Academy of Sciences 49(6): 871-878.
Seo, H. Y., J. M. Park, et al. (2010). "Prognostic significance of serum vascular endothelial growth factor per platelet count in unresectable advanced gastric cancer patients."
Japanese Journal of Clinical Oncology 40(12): 1147-1153.
Solter, D. and B. B. Knowles (1978). "Monoclonal antibody defining a stage-specific mouse embryonic antigen (SSEA-1)." Proc Natl Acad Sci U S A 75(11): 5565-5569.
Sulpice, E., J. Plouet, et al. (2008). "Neuropilin-1 and neuropilin-2 act as coreceptors, potentiating proangiogenic activity." Blood 111(4): 2036-2045.
Sulzenbacher, G., C. Bignon, et al. (2004). "Crystal structure of Thermotoga maritima alpha-L-fucosidase. Insights into the catalytic mechanism and the molecular basis for fucosidosis." J Biol Chem 279(13): 13119-13128.
Sun, Y., K. Jin, et al. (2004). "Increased Severity of Cerebral Ischemic Injury in Vascular Endothelial Growth Factor-B-Deficient Mice." J Cereb Blood Flow Metab 24(10):
1146-1152.
Takahashi, H., T. Saibara, et al. (1994). "Serum alpha-L-fucosidase activity and tumor size in hepatocellular carcinoma." Hepatology 19(6): 1414-1417.
Takeshita, S., L. P. Zheng, et al. (1994). "Therapeutic angiogenesis. A single intraarterial bolus of vascular endothelial growth factor augments revascularization in a rabbit ischemic hind limb model." J Clin Invest 93(2): 662-670.
Terentiev, A. and N. Moldogazieva (2006). "Structural and functional mapping of α-fetoprotein." Biochemistry (Moscow) 71(2): 120-132.
Tomasi, T. B., Jr. (1977). "Structure and function of alpha-fetoprotein." Annu Rev Med 28:
453-465.
Tseng, P.-L., M.-H. Tai, et al. (2008). "Overexpression of VEGF is associated with positive p53 immunostaining in hepatocellular carcinoma (HCC) and adverse outcome of HCC patients." Journal of Surgical Oncology 98(5): 349-357.
Unemori, E. N., N. Ferrara, et al. (1992). "Vascular endothelial growth factor induces interstitial collagenase expression in human endothelial cells." J Cell Physiol 153(3):
557-562.
VESTWEBER, D. and J. E. BLANKS (1999). "Mechanisms That Regulate the Function of the Selectins and Their Ligands." Physiological Reviews 79(1): 181-213.
Wada, H., H. Nagano, et al. (2006). "Expression pattern of angiogenic factors and prognosis after hepatic resection in hepatocellular carcinoma: importance of angiopoietin-2 and hypoxia-induced factor-1a." Liver International 26(4): 414-423.
Wang, L., H. Zeng, et al. (2003). "Neuropilin-1-mediated vascular permeability factor/vascular endothelial growth factor-dependent endothelial cell migration." J Biol Chem 278(49): 48848-48860.
Wu, J. T. and J. A. Knight (1985). "In-vitro stability of human alpha-fetoprotein." Clinical Chemistry 31(10): 1692-1697.
Wu, J. T. and W. J. Waterhouse (1982). "Identification of alpha-fetoprotein polymers. Artifacts of the isolation procedure." Clinica Chimica Acta 125(1): 9-19.
Xu, X. and E. R. Kantrowitz (1991). "A water-mediated salt link in the catalytic site of Escherichia coli alkaline phosphatase may influence activity." Biochemistry 30(31):
7789-7796.
Yao, D. F., X. H. Wu, et al. (2005). "Quantitative analysis of vascular endothelial growth factor, microvascular density and their clinicopathologic features in human hepatocellular carcinoma." Hepatobiliary Pancreat Dis Int 4(2): 220-226.
Zhang, F., Z. Tang, et al. (2009). "VEGF-B is dispensable for blood vessel growth but critical
for their survival, and VEGF-B targeting inhibits pathological angiogenesis."
Proceedings of the National Academy of Sciences 106(15): 6152-6157.
Zhou, L., J. Liu, et al. (2006). "Serum tumor markers for detection of hepatocellular carcinoma." World J Gastroenterol 12(8): 1175-1181.
Zinkin, N. T., F. Grall, et al. (2008). "Serum proteomics and biomarkers in hepatocellular carcinoma and chronic liver disease." Clin Cancer Res 14(2): 470-477.