Development of an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. A molecular biomarker for cataract.

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dc.contributor.authorNayak, Sujathaen_US
dc.contributor.authorSashidhar, R Ben_US
dc.contributor.authorBhat, K Seetharamen_US
dc.date.accessioned2002-03-02en_US
dc.date.accessioned2009-05-29T10:21:46Z
dc.date.available2002-03-02en_US
dc.date.available2009-05-29T10:21:46Z
dc.date.issued2002-03-02en_US
dc.description.abstractPURPOSE: To develop and evaluate an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. MATERIALS AND METHODS: Beta and gamma-crystallins isolated from rat lens were used as immunogens to raise polyclonal antibodies in rabbits. Antibody capture assay and western blot analysis showed that the antibodies to beta- and gamma-crystallins were specific. An indirect competitive enzyme linked immunosorbent assay (ELISA) developed to quantitate beta- and gamma-crystallin showed an IC50 value of 70 ng and 65 ng, respectively, based on regression analysis. Spiking studies with purified beta-crystallin antibodies showed that 33 ng of the purified antibody gave an absorbance of 1.1 at 450 nm, indicating the sensitivity of the method. RESULTS: Antibodies to beta- and gamma-crystallins were not detected in serum samples of the cataractous CFY/NIN rats (used as an animal model for induction of experimental cataract by feeding high galactose diet). However, the cataractous rat serum samples effectively displaced beta- and gamma-crystallin antibodies, indicating that these crystallins leak during cataract formation. The concentration of beta- and gamma-crystallins in the rat serum, as analysed by indirect competitive ELISA, was found to be in the range of 17.6-81.6 micrograms/ml [corrected] and 12.4-19.6 micrograms/ml, respectively. CONCLUSIONS: The methodology developed in the present study may find application as a biochemical tool in molecular epidemiology of cataract.en_US
dc.description.affiliationDepartment of Biochemistry, University College of Science, Osmania University, Hyderabad-500 007, India.en_US
dc.identifier.citationNayak S, Sashidhar RB, Bhat KS. Development of an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. A molecular biomarker for cataract. Indian Journal of Ophthalmology. 2002 Mar; 50(1): 41-8en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/72601
dc.language.isoengen_US
dc.source.urihttps://www.ijo.inen_US
dc.subject.meshAnimalsen_US
dc.subject.meshAutoantibodies --blooden_US
dc.subject.meshBiological Markers --analysisen_US
dc.subject.meshBlotting, Westernen_US
dc.subject.meshCataract --diagnosisen_US
dc.subject.meshCrystallins --immunologyen_US
dc.subject.meshElectrophoresis, Polyacrylamide Gelen_US
dc.subject.meshEnzyme-Linked Immunosorbent Assay --methodsen_US
dc.subject.meshLens, Crystalline --chemistryen_US
dc.subject.meshMaleen_US
dc.subject.meshModels, Animalen_US
dc.subject.meshRabbitsen_US
dc.subject.meshRatsen_US
dc.subject.meshRats, Wistaren_US
dc.titleDevelopment of an immunoanalytical method for the detection of beta- and gamma-crystallins and anti-crystallin antibodies. A molecular biomarker for cataract.en_US
dc.typeJournal Articleen_US
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