I'm working on a lateral flow immunoassay. Conjugated to my colloidal gold is goat anti-bovine IgG. For the control line, I immobilized bovine IgG. My boss suggested to use anti-goat IgG to capture the conjugate instead. I have read some journals doing such as well wherein they used the antibody against the conjugate protein in the control line. Is it any better than using the protein (i.e. bovine IgG) the conjugate antibody (goat anti-bovine IgG) is against for in the control line? If so, how and why?
From my point of view, the use of bovine IgG as a control line is fine. However, you should wash the line properly to avoid any soluble bovine IgG to be present. The use of anti-goat antibody is also possible. It may have the disadvantage that even an inactive conjugate leads to a positive control line.
Thank you Sir for the insightful answer! It seems then that bovine IgG is the better choice as it additionally tests for the functionality of the conjugate. I think I'm gonna go with bovine IgG.
Stjepan Krešimir Kračun
If I will use polyclonal anti-bovine IgG in my control line, then how will the conjugate be captured in the line? Indirectly via bovine IgGs in the sample?
Stjepan Krešimir Kračun
Thank you for the clarification! But do you mean polyclonal anti-goat IgG? My conjugate protein is anti-bovine IgG from goat.
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