Creating and unveiling chiral systems is important to the development of new materials and devices. In this study, after depositing prochiral radical molecule 3-carbamoyl-2,2,5,5-tetramethyl pyrroline-1-oxyl (CTPO) on a Au(111) substrate, 2D molecular crystals with two chiralities of CTPO molecules have been discovered. A single CTPO molecule is an achiral molecule in three-dimensional space, and it can form chiral configurations after adsorbing on the substrate. The chiralities of 2D molecular crystals originate from the chiral properties of adsorbed CTPO molecules and their assembling. Molecules with different chiralities are connected with molecular recognition through hydrogen bond interactions. Through density functional theory simulations, the hydrogen bond networks and molecular structures of this system were explored. To gain a further understanding of this system, the electronic property of CTPO/Au(111) was studied with a local density of states (LDOS) characterization. A peculiar LDOS distribution related to the vibrational excitation of the molecules was mapped at the submolecular scale. These results are useful for understanding the nature of chirality formation, 2D molecular crystal construction, and radical molecule applications.