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Use of Heavy Water (D2O) in Developing Thermostable Recombinant p26 Protein Based Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Equine Infectious Anemia Virus Infection

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Title Use of Heavy Water (D2O) in Developing Thermostable Recombinant p26 Protein Based Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Equine Infectious Anemia Virus Infection
Not Available
 
Creator Harisankar Singha
Sachin K. Goyal
Praveen Malik
Raj K. Singh
 
Subject Equine infectious anemia
recombinant p26
Heavy Water
ELISA
Thermostabilization
 
Description Not Available
Thermostabilizing effect of heavy water (D2O) or deuterium oxide has been demonstrated previously on several enzymes and
vaccines like oral poliovirus vaccine and influenza virus vaccine. In view of the above observations, effect of heavy water on in
situ thermostabilization of recombinant p26 protein on enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of equine
infectious anemia virus (EIAV) infection was investigated in the present study. The carbonate-bicarbonate coating buffer was
prepared in 60% and 80% D2O for coating the p26 protein in 96-well ELISA plate and thermal stability was examined at 4โˆ˜C,
37โˆ˜C, 42โˆ˜C, and 45โˆ˜Cover a storage time from2weeks to 10months. A set of positive serum (๐‘› = 12) consisting of strong, medium,
and weak titer strength (4 samples in each category) and negative serum (๐‘› = 30) were assessed in ELISA during the study period.
At each time point, ELISA results were compared with fresh plate to assess thermal protective effect of D2O. Gradual increase in
the stabilizing effect of 80% D2O at elevated temperature (37โˆ˜C < 42โˆ˜C < 45โˆ˜C) was observed. The 80% D2O provides the thermal
protection to rp26 protein in ELISA plate up to 2 months of incubation at 45โˆ˜C. The findings of the present study have the future
implication of adopting cost effective strategies for generating more heat tolerable ELISA reagents with extended shelf life.
Not Available
 
Date 2020-07-31T05:50:27Z
2020-07-31T05:50:27Z
2014-01-09
 
Type Article
 
Identifier Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/38632
 
Language English
 
Publisher Hindwai