Detection Ab - Biotinylated and Not Biotinylated

Dear SPR-pages,

We have a customer that wants to test 2 Ab's that will be Detection Ab's. Determining if they are a match pair with the capture Ab. One will be biotinylated while the other one Will Not be biotinylated. We have this method set up (see photo attached) that has worked pretty well. However, all the Ab's we have tested to date is NOT biotinylated. They wanted to know if biotinylation of the Detection Ab will affect its' binding characteristics. Will this set up work? Hope this makes sense. Thanks.

If you label proteins generally the label will be attached ad random depending on the type of labelling procedure used. Since labelling is ad random a fraction of the labelled IgG will be not functional anymore because the label interferes with binding. You can easily test this with SPR by injection of unlabeled and labeled IgG and look at the differences.

Kinds regards
Arnoud