2 replies to this topic

[syloc]

Hi, i am new here.

I have an enquiry. What type of recombinant protein is suitable as antigen for ELISA? What i meant was soluble or inclusion body protein? I been trying to find the answer but fail. Does anyone know if inclusion body was good for ELISA? I am currently trying to express 7 proteins as antigen to detect antibody in patients sera. I manage to obtain inclusion body protein but was having a problem in obtaining the soluble fractions. I don't want to waste much of my time in optimizing the expression as it is not the aim of my project. Can anyone advice me on this? If anyone has any reference regarding this, I would be greatly happy if you can share it with me. Thanks!!

[sgt4boston]

found this link/article

http://www.sciencedirect.com/science?_ob=A...940e9aaba2613e7

Purification of the extra-cellular domain of Nipah virus glycoprotein produced in Escherichia coli and possible application in diagnosis

Majid Eshaghia, Wen Siang Tana, b, Wai Kit China and Khatijah Yusoffa, b,


Hope it is helpful.

[Raj2009]

Dear D,

It is good to have recombinant protein expressed as soluble form when you are using it to develope antibodies and then ELISA.

Because soluble form of antigen as following advantages:

Easy to get specific antibodies as there is no cross contaminating proteins.
When you want to develop quantitative or semi quantitative ELISA you can use soluble recombinant protein as standard positive controls. It is convenient to have different concentrations of calibrators.