Indian Journal of Ophthalmology

ORIGINAL ARTICLE
Year
: 2002  |  Volume : 50  |  Issue : 1  |  Page : 41--48

Development of an immunoanalytical method for the detection of β- and γ- Crystallins and anti-crystallin antibodies. A molecular biomarker for cataract


S Nayak, RB Sashidhar, K Seetharam Bhat 
 Department of Biochemistry, University College of Science, Osmania University, Hyderabad-500 007, India

Correspondence Address:
S Nayak
Department of Biochemistry, University College of Science, Osmania University, Hyderabad-500 007
India

Purpose: To develop and evaluate an immunoanalytical method for the detection of β - and g-crystallins and anti-crystallin antibodies. Materials and Methods: Beta and g-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 β - and g-crystallins were specific. An indirect competitive enzyme linked immunosorbent assay (ELISA) developed to quantitate β - and g-crystallin showed an IC50 value of 70 ng and 65 ng, respectively, based on regression analysis. Spiking studies with purified β -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 β - and g-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 β - and g-crystallin antibodies, indicating that these crystallins leak during cataract formation. The concentration of β - and g-crystallins in the rat serum, as analysed by indirect competitive ELISA, was found to be in the range of 17.6 - 81.6 mg/l and 12.4- 19.6 g/ml, respectively. Conclusions: The methodology developed in the present study may find application as a biochemical tool in molecular epidemiology of cataract


How to cite this article:
Nayak S, Sashidhar R B, Bhat K S. Development of an immunoanalytical method for the detection of β- and γ- Crystallins and anti-crystallin antibodies. A molecular biomarker for cataract.Indian J Ophthalmol 2002;50:41-48


How to cite this URL:
Nayak S, Sashidhar R B, Bhat K S. Development of an immunoanalytical method for the detection of β- and γ- Crystallins and anti-crystallin antibodies. A molecular biomarker for cataract. Indian J Ophthalmol [serial online] 2002 [cited 2021 Jan 23 ];50:41-48
Available from: https://www.ijo.in/article.asp?issn=0301-4738;year=2002;volume=50;issue=1;spage=41;epage=48;aulast=Nayak;type=0