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Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.48 [2010] >
IJEB Vol.48(08) [August 2010] >
| Title: | Preparation and characterization of immunoglobulin yolk against the venom of Naja naja atra |
| Authors: | Liu, Sihong
Dong, Weihua
Kong, Tianhan |
| Keywords: | Egg yolk
Immunoglobulin
Naja naja atra
Snake venom |
| Issue Date: | Aug-2010 |
| Publisher: | CSIR |
| Abstract: |
Chinese Cobra (Naja naja atra) bite is one of the
leading causes of snake-bite mortality in China. The traditional anti-cobra
venom serum therapy was found to be expensive and with high frequency of side
effects. Therefore attempts were made to generate a high titer immunoglobulin from egg yolk (IgY) of crude
cobra-venom immunized Leghorn
hens, and to standardize an effective method for producing avian antivenom in
relatively pure form. The IgY was isolated first by water dilution method to
remove the lipid, then extracted by ammonium sulfate precipitation, and
purified through anion exchange chromatogram. The different purities of IgY
from different isolating stages were submitted to enzyme-linked immunosorbent
assay and SDS-PAGE to determine their titers. Immunoblotting showed that the
purified IgY (ion exchange chromatography fraction, IECF) recognized several
antigenic fractions of cobra venom, and presented with the character of
polyclonal antibody. IECF on SDS-PAGE under reducing conditions migrated as a
65 kDa heavy chain and a 35 kDa light chain, respectively. The LD50
of the N. naja atra venom was 0.62 mg/kg body weight in mice. Four times
the LD50 dose of venom was selected as challenge dose,
and the ED50 of IgY was 3.04 mg IECF/mg venom. The results indicate
that the activity of anti-snake venom IgY could be obviously elevated by ion
exchange chromatography, thus possessing therapeutic significance for snakebite
envenomation.
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| Page(s): | 778-785 |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.48(08) [August 2010]
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