Oriented immobilization of antibodies using boronic acids shows a strong potential for improving immunoassay performance but is not yet widely used, possibly because of the difficulties encountered in its implementation. How to choose the boronic acid structure and how should it be attached to the surface? How to choose an antibody that will bind to the boronic acid? Under which conditions should the binding take place for an effective oriented antibody immobilization? How to make sure that the antibody stays on the surface? This tutorial review provides answers to these questions through analysis of the literature and personal suggestions, and thereby intends to facilitate the development of this promising antibody immobilization strategy.
A new method for the selective and sensitive detection of boronic acids on thin-layer chromatography plates is described. The plate is briefly dipped in an alizarin solution, allowed to dry in ambient air, and observed under 366 nm light. Alizarin emits a bright yellow fluorescence only in the presence of a boronic acid.
Regioselective O-alkylation of indazolinones using (cyanomethylene)triphenthylphosphorane (CMPP) as a Mitsunobutype reagent is described with a variety of aliphatic alcohols. This method was also successfully applied to the N-alkylation of O-protected indazolinone. Selective N1 and N2 alkylations on indazolinone have previously been described; our methodology is therefore orthogonal to the previous precedent.
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