Prasad, Pramod Vishwanath and Rani, Aruna and Chaube, Shail Kumar and Rohil, Vishwajeet and Shrivastav, Tulsidas G (2008) Kinetic analysis of a human chorionic gonadotropin- epitope-paratope interaction. In: Growth Factors, 26 (6). pp. 331-342.
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Kinetics of protein-protein or ligand-ligate interaction has predominantly been studied by optical spectroscopy (particularly fluorescence) and surface plasmon resonance biosensors. Almost all such studies are based on association kinetics between ligand-ligate and suffer from certain methodological and interpretational limitations. Therefore, kinetic analyses of dissociation data of such interactions become indispensable. In the present investigation, the radiolabeled human chorionic gonadotropin- (125IhCG) was employed as a probe and nitrocellulose (NC) as a solid support to immobilize monoclonal antibody (MAb) G1G10.1. The NC-G1G10.1-125IhCG complex (NCcom) was prepared and the dissociation of radiolabeled hCG was carried out in the presence of excess unlabeled ligate. From the experimental dissociation data under varying ionic strength, dissociation constants (k-1), association constants (k+1) and affinity constants (ka) were calculated. The values obtained were utilized in exploring the amino acid residues constituting an epitopic region of hCG involved in interaction with the complementary paratope on MAb G1G10.1. Kinetic data of the present study supported our recently published findings [using single step-solid phase radioimmunoassay (SS-SPRIA)] that the core region of hCG epitope consists of Arg (94,95) and Asp (99) while a Lys (104) and a His (106) are in proximity to the core epitopic region. Based on the results of present investigation, we conclude that dissociation kinetics coupled with SS-SPRIA unequivocally provides considerable insight into the study of ligand-ligate interactions and epitope analysis.
| Item Type: | Journal Article |
|---|---|
| Publication: | Growth Factors |
| Publisher: | Taylor and Francis Group |
| Additional Information: | Copyright of this article belongs to Taylor and Francis Group. |
| Keywords: | Human chorionic gonadotropin -subunit;MAb G1G10;1;antigen antibody;kinetic constants. |
| Department/Centre: | Division of Biological Sciences > Molecular Reproduction, Development & Genetics |
| Date Deposited: | 27 Aug 2009 06:14 |
| Last Modified: | 27 Aug 2009 06:14 |
| URI: | http://eprints.iisc.ac.in/id/eprint/17841 |
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