Herein we present a proof-of-concept study of target-dependent gating of nanopores for general photoelectrochemical (PEC) bioanalysis in an H-cell. The model system was constructed upon a left chamber containing ascorbic acid (AA), the antibody modified porous anodic alumina (AAO) membrane separator, and a right chamber placed with the three-electrode system. The sandwich immunocomplexation and the associated enzymatic generation of biocatalytic precipitation (BCP) in the AAO nanopores would regulate the diffusion of AA from the left cell to the right cell, leading to a varied photocurrent response of the ZnInS nanoflakes photoelectrode. Exemplified by fatty-acid-banding protein (FABP) as the target, the as-developed protocol achieved good performance in terms of sensitivity, selectivity, reproducibility, as well as efficient reutilization of the working electrode. On the basis of an H-cell, this work features a new protocol of target-dependent gating-based PEC bioanalysis, which can serve as a general PEC analytical platform for various other targets of interest.