To provide a method of manufacturing a biochip of high sensitivity as high throughput constituted so as to fix a physiologically active material such as protein, a molecule for capturing it or the like to the surface of a substrate at an arbitrary position but not to adsorb an unnecessary physiologically active material to a part other than the arbitrary position and capable of detecting the physiologically active material.

The method of manufacturing the biochip constituted by fixing a capture being a material, which captures the physiologically active material specifically, to the surface of the substrate in a spot-like state includes a step (1) for preliminarily covering the part, where the capture is fixed, of the surface of the substrate with a masking material, a step (2) for coating the surface of the substrate, which includes the part covered with the masking material, with a material for suppressing the bonding or absorption of the plysiologically active material, a step (3) for removing the masking material and a process (4) for bringing a capture-containing solution into contact with the part where the masking material is removed.