<p>Chirality plays a critical role in the structure and function of natural and synthetic polymers, impacting their mechanical, optical, and electronic properties. However, a comprehensive understanding of the hierarchical emergence of chirality from monomers to macromolecular assemblies remains elusive, largely because of current limitations in studying their chemical-structural properties at the nanoscale. Here, we unravel the emergence of different forms of chirality from small molecules to their resulting polymers and supramolecular assemblies. We leverage bulk spectroscopic methods combined with the development of acoustical-mechanical suppressed infrared nanospectroscopy, to empower chemical-structural analysis of single-polymer chains. This ultra-high sensitivity allows identifying key functional groups as a signature for different forms of chirality: CH groups for central chirality in small molecules; C=O groups for backbone and supramolecular chirality in heterogeneous polymers. This work opens a new single-molecule chemical angle of observation into chirality and (bio)-polymers for the rational design in material science, biotechnology, and medicine.</p>