Home-made Hepatitis B Surface Antigen

S. Saengamnatdej, Ph.D.
August 19, 2010

Our pure positive antigen for testing hepatitis B viral infection was running out
and I need certain amount of it in a next couple of days. We knew that we had some
peripheral blood donated from a carrier in the University Hospital kept in a -20 °C
freezer.
We therefore tried making the antigen from the blood. The protocol that we used
was very easy and the antigenic property of the antigen was tested.

Protocol
A. Preparation of HBsAg
1. Thaw the frozen blood completely at room temperature.
2. Heat it in boiling water for 5 minutes.
3. Remove it from the water bath and leave it to cool down a bit.
4. Spin in a centrifuge at 3000xg for 5 minutes.
5. Decant the clear solution into new eppendorfs

B. Testing the antigen

1. Apply 50 microlitres of the antigen (step 5, above) onto the pad of a test strip
for detection of HBsAg.
2. Wait for 5 minutes and read the result.

C. Result
There were two bands on the test strip; one was the control (the
upper one) and the other was the test band (see the picture on the
right) of the current antigen.

D. Discussion and Conclusion

The antigen was intended to be used in the demonstration and
training a student in a course, so we needed to make very sure that the
virus and all those infectious or non-infectious bio-hazards in the blood
were completely inactivated. This would make the handle of the
antigen much easier. The blood cells and proteins turned to pale gray
which suggested that they were denatured and had lost their native
biological activities during the heating step. The surface protein of the
hepatitis B virus would be denatured in some level as well, but
fortunately, as tested, its antigenic epitope was intact, and could be
bound and immobilized by the anti-HBsAg in the strip.
We therefore concluded that this method is good enough to prepare the HBsAg.