I am currently creating a direct ELISA to measure an HIV protein in a viral vector. I'm using the formulated buffer to generate my standard curve, and it works beautifully. However, when I do spike recovery experiments in the vector itself, I only recover approximately 10-20% of my input. I did a dilution series of vector to buffer down to 1:2000, and I continue to get the same OD values no matter the dilution. Does anyone have experience with this? I've never done an ELISA with a viral vector, so this step is new to me. I know the vector is interfering, but based on my results, I don't know how. Any advice at all would be greatly appreciated!
Hello Dactor, yes must be depending because sensitive and specific between variate kits.
Jessica, If I understand right when you use the viral vector in the dilution series instead of the recombinant protein, After the dilution range of 1:2000, you are able to see same OD reading?
Concerns
1) How did you differentiate between free HIV protein and Virus associated protein
2) Check if your sample dilution buffer is contaminated
3) ELISA is sensitivity to range of the standards, please check them
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