Ayusoff, N. and Soin, N. and Abd Rahman, N. and Raja Abd Rahman, R.N.Z. and Abd Kahar, M.K.B. and Ibrahim, F. (2009) Binding characteristics study for dengue virus non-structural protein 1 of antigen and Its antibody by using circular dichroism technique. In: International Conference for Technical Postgraduates 2009, TECHPOS 2009, 2009, Kuala Lumpur.
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Abstract
This paper presents the binding characteristics study of dengue non-structural protein 1 (NS1) antigen and its antibody using circular dichroism technique in far UV region. Circular dichroism (CD) is a spectroscopic technique which measures differences in the absorption of left-handed and right handed circularly polarized light. The CD spectrum can determine conformation of the NS1 antigen and its antibody, conformational changes of the antigen-antibody interaction and estimates the secondary structure of these proteins in far UV region. Firstly, CD spectrum of individual solutions of the antigen and the antibody were measured. Then, the solutions were mixed to produce a solution of complex dengue NS1 antigen and its antibody for measurement. The findings show that the antibody has the highest positive band of CD intensity follow by the complex antigen-antibody and antigen. The antibody is a chiral structure, has high helical conformation and more ordered epitope structure. Meanwhile, the NS1 antigen shows the negative and the lowest CD spectrum. The antigen is low chirality and has more random-like conformation. The complex (binding of the antigen and antibody) has the CD spectrum's shape similar to the antibody but in lower intensity. So, it has helical and beta conformations lower than the antibody. The binding characteristics of the complex solutions were also studied with increased in incubation time and with varied rotation applied. It is found that the immunoreactions between the antigen and its antibody are rapid processes which do not require too long incubation time. Besides, the applied rotation can increased the immunoreaction process.
| Item Type: | Conference or Workshop Item (Paper) |
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| Funders: | UNSPECIFIED |
| Additional Information: | Conference code: 79876 Export Date: 29 January 2014 Source: Scopus Art. No.: 5412060 doi: 10.1109/TECHPOS.2009.5412060 Language of Original Document: English Correspondence Address: Ayusoff, N.; Dept. of Biomedical Engineering, Faculty of Engineering, University Malaya, K.LMalaysia; email: aizas05@yahoo.com References: Tinoco Jr., I., Sauer, K., Wang, J.C., Puglisi, J.D., (2002) Physical Chemistry. Principle and Applications in Biological Sciences, pp. 573-575. , 4th, Prentice Hall; Kelly, S.M., Jess, T.J., Price, N.C., How to study proteins by circular dichroism (2005) Biochimia et Biophysica Acta, 1751, pp. 119-139. , July; (2009) Ministry of Health Estimates Less Than 1000 Dengue Cases per Year, , http://htaiping.moh.gov.mv/modules/news/article.php?storvid=18, IT Unit Taiping Hospital. Online. April 14 cited 2009 Aug 28; Mackenzie, J.M., Jones, M.X., Young, P.R., Immunolocalization of the Dengue virus nonstructural glycoprotein NS1 suggests a role in viral RNA replication (1996) Virology, 220, pp. 232-240; Rossier, J.S., Gokulrangan, G., Girault, H.H., Svojanovsky, S., Wilson, G.S., Characterization of protein adsorption and immunosorption kinetics in photoablated polymer microchannels (2000) Langmuir: The ACS Journal of Surfaces and Colloids, 16 (22), pp. 8489-8494; Lai, S., Wang, S., Luo, J., Lee, J., Yang, S.T., Madou, M.J., Design of a compact disk-like microfluidic platform for enzyme-linked immunosorbent assay (2004) Annal. Chemistry, 76, pp. 1832-1837. , April; (2009) Dengue Virus NS1 Glycoprotein Protein (Ab64456), , http://www.abcam.com/Dengue-Virus-NS1-glycoprotein-proteinab64456.html, Abeam plc. Product Documentation. Online, cited Aug 28; Dengue Virus NS1 Glycoprotein Antibody DN3 (Ab41616), , http://www.abcam.com/Dengue-Virus-NS1.-glycoproteinantibody-DN3-ab41616. html, Abeam plc. Product Documentation. Online, cited 2009 Aug 28; Sreerama, N., Yu Venyaminov, S., Woody, R.W., Estimation of protein secondary structure from circular dichroism spectra: Inclusion of denatured proteins with Native proteins in the analysis (2000) Analytical Biochemistry, 287, pp. 243-251. , April; Douglas, J., (2009) An Introduction, to Circular Dichroism, , http://mass-spec.chem.cmu.edu/procedurefiles/cd/cdeuide.doc, Online, cited Aug 25; Laczko, I., Bottka, S., Toth, G.K., Malvy, C., Bertrand, J.-R., Hollosi, M., Interaction of fusogenic peptides with an antisense oligonucleotide in solution and in the presence of micelles: Conformational studies (2003) Biochemical and Biophysical Research Communications, 313 (2004), pp. 356-361. , November; Mattu, T., Pleass, R., Willis, A., Kilian, M., Wormald, M., Lellouch, A., Rudd, P., Dwek, R., The glycosylation and structure of human, serum. IgA1, Fab, and Fc regions and the role of Nglycosylation on Fc alpha receptor interactions (1998) J Biol Chem, 273 (4), pp. 2260-2272; Wallis, T.P., Huang, C.Y., Nimkar, S.B., Young, P.R., Gorman, J.J., Determination of the disulfide bond arrangement of dengue virus NS1 protein (2004) J. Biol. Chem., 279, pp. 20729-20741; Flamand, M., Megret, F., Mathieu, M., Lepault, J., Rey, F.A., Deubel, V., Dengue Virus type 1 nonstructural glycoprotein. NS1 is secreted from mammalian cells as a soluble hexamer in a glycosylation-dependent fashion (1999) J. Of. Virology, 73 (7), pp. 6104-6110. , July |
| Uncontrolled Keywords: | Antigen-antibody interaction, Binding characteristics, CD spectra, Chiral structures, Circular dichroism, Circularly polarized light, Complex solution, Conformational change, Dengue virus, Helical conformation, Incubation time, Right handed, Secondary structures, Spectroscopic technique, Structural proteins, UV region, Absorption spectroscopy, Antigens, Complexation, Conformations, Data storage equipment, Dichroism, Rotation, Stereochemistry, Viruses, Antibodies |
| Subjects: | T Technology > T Technology (General) T Technology > TA Engineering (General). Civil engineering (General) |
| Divisions: | Faculty of Engineering |
| Depositing User: | Mr Jenal S |
| Date Deposited: | 17 Feb 2014 08:27 |
| Last Modified: | 01 Nov 2017 05:28 |
| URI: | http://eprints.um.edu.my/id/eprint/9264 |
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