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The Profile of the Expression of IBMR3 Antigens 3T3 and HT29 Cancer Cell Line after Analysis
Science Research
Volume 2, Issue 4, August 2014, Pages: 55-61
Received: Jun. 30, 2014; Accepted: Jul. 15, 2014; Published: Jul. 30, 2014
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Qutaiba Kafi Jassim Alrawi, Laboratory Department, faculty of Medicine Technology of Surman, Alzawiah University, Surman, Libya
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Aims: The aim of this study was to evaluate the effectiveness of specific monoclonal IBMR3 antibodies expression in the cancer cell lines of 3T3 and HT29, in order to recognize specific antigen and make evaluation, using immunoblotting technique which is widely use in this filed. Methods: Protein extracts were extracted from these cancer cell lines and used for western bloat. The Immunobloting were consequently subjected to densitometry analysis using bioimaging machine. This bioimaging process will facilitate to measure the molecular weights, peak height and raw volume of the protein bands for the 3T3 and HT29 cancer cell lines, which helps in diagnosis of any pathogenic antigen. Results: The bands obtained from bioimaging were exposed on the PVDF membrane. In the 3T3 bio imaging process revealed four bands and molecular weight were 299.58, 87.90, 41.67 and 23.54 KDa. However, bioimaging results for HT29 revealed also four bands with molecular weights: 90.11, 41.31, 23.87 and 20.86 KDa. The results of Peak height densitometry for IBMR3 antigen bands for 3T3 were: 1856.985, 551.769, 394.164, and 216.185. However HT29 peak height results were: 281.544, 101.711.202.668, 757.213. The raw volume (amount of protein bands of IBMR3 Ag for 3T3) were: 1460168.75, 206078.47, 161406.89, and 219583.16. However HT29 raw volume results were: 110197.11, 76106.84, 98632.59, and 221395.34. Negative protein staining for 3T3 and HT29 cancer cell line bands were done by using mouse IgM serotype. IgM serotype was not indicated that means no specific antigens for negative control IgM. Conclusion: The bioimaging revealed different results of the expression profile in molecular weight, peak height and raw volume between the two cancer cell lines. The results from this study suggest that the IBMR3 antigens were differentially expressed in 3T3 and HT29 and the molecular weight is higher in the 3T3 than HT29 cancer cell lines. In future it will be beneficial to categorize and study the character analysis of the IBMR3 antigen and its prospective role in cancer cells.
3T3 (Mouse Fibroblast) Cell Line and HT29 (Human Colorectal) Carcinoma Cancer Cell Line, SDS-Page,Monocloncal Antibodies, MAb IBMR3, Immunobloting, Bioimaging
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Qutaiba Kafi Jassim Alrawi, The Profile of the Expression of IBMR3 Antigens 3T3 and HT29 Cancer Cell Line after Analysis, Science Research. Vol. 2, No. 4, 2014, pp. 55-61. doi: 10.11648/
Boersma, W. J. A., Claassen, E., Deen, C., Gerritse, K., Haaijman, J. J. and Zegers, N. D. (1988) Antibodies to short synthetic peptides for specific recognition of partly denatured protein. Analytical chemistry Acta, 213, 187- 197.
Galizzi, J. P., Zuber, C.E., Cabrillat, H., Gorman, M., Djossou, O., Kastelin, R.and Banchereau, J. (1990) Molecular cloning of a cDNA encoding the human interleukin-4 receptor. International Immunology, 2, 669-675.
Hara, Y and Mat, I.B. (2004) Differential expression of IBMR3 antigens in normal and transformed cells. Medimond International Proceedings: Immunology, E7 18C4844, 229- 233.
Hawkins, R.E., Llewelyn, M.B. and Russell, S.J. (1992) Adapting antibodies clinical use. Biomedical Journal, 305, 1348-1352.
Kaspar, R. (1997) Western Blotting (a laboratory protocol).
Kinoshita, T., Inoue, N. (2000) Dissecting and manipulating the pathwayfor glycosylphosphatidylinositol-anchor biosynthesis. Curr Opin Chem Biol, 4,632–638.
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680-685.
Molina, H., Holers, V. M., LI, B., Fang, Y-F., Mariathsan, S., Goellner, J., Strauss-Schoenberger, J., Karri, R.W., and Chaplin, A. D. (1996) Markedly impaired humoral immune response in mice deficient in complement receptors 1 and 2. Proceedings National Academy of Sciences of the United States of America, (93), 3357-3361.
Mat, I.B. (1992) Analysis of human interleukin-4 receptor-associated molecules (gp200-MR6 molecule) in normal and transformed epithelia. PhD thesis, London: University of London, 244- 277.
Murillo, O., Arina, A., Tirapu, I., Alfaro, C., Mazzolini, G., Palencia, B., López-Diaz De Cerio, A., Prieto, J., Bendandi, M. and Melero, I. (2003) Potentiation of Therapeutic Immune Responses against Malignancies with Monoclonal Antibodies. Clinical Cancer Research. (9), 5454-5464.
Paulick, M.G., Bertozzi, CR. (2008) the glycosylphosphatidylinositol anchor: A complex membrane-anchoring structure for proteins. Biochemistry, 47, 6991–7000.
Reichert, J., M. and Valge-Archer, V., A. (2007) Outlook: Development trends for monoclonal antibody cancer therapeutics, Nature Reviews Drug Discovery, 6, 349-356.
Shapiro, A.L., Vinuela, E.and Maizel, J.V. Jr. (1967) Molecular Weight Estimation of Polypeptide Chains by Electrophoresis in SDS Polyacrylamide Gels. Biochemical and Biophysical Research Communications, 28(5), 815-820.
Stryer, L. (1996) Biochemistry. New York: W.H. Freeman, 62.
Spinks, C.A. (2000). Broad-specificity immunoassay of low molecular Weight food contaminants new paths to Utopia Trend. Food Science and Technology, 11, 210–217.
Towbin, H., Staehekin, T. and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings National Academy of Sciences of the United States of America, 76(9), 4350-4354.
Walter, G. and Doolittle. (1983) Antibodies against synthetic peptides. Genetic Engineering, 61-6
Abbreviations: mAb (monoclonal anti body), HRP (horse radish peroxidase), PVDF (Polyvinylidene difluoride membrane), IgG (Immunoglobulin G), IgM (Immunoglobulin M), (IgS Immunoglobulin S). IL-4 (Human Interleukin – 4. recapture)
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