A method for manufacturing a glass-based biosensor is used to solve the problem of the use of a solution containing a strong acid or a strong base or of an oxygen plasma treatment. The method comprises modifying a silicon-containing substrate by an alcohol solution to form negative charges on at least one coupling surface of the silicon-containing substrate. A least one active layer of polymer having positive charges is formed on the at least one surface of the silicon-containing substrate, respectively. Each of the at least one active layer of polymer has a coupling surface and an active surface opposite to the coupling surface, and the at least one active layer of polymer couples to the silicon-containing substrate via the coupling surface. A plurality of capture biomolecules couples to the active surface.