The invention discloses a method for large-scale separation and purification of recombinant human-derived collagen. The space radius of protein molecules is accurately controlled by conducting primary folding and refolding on linear recombinant protein, convenience is brought to accurate aperture selection of microfiltration and ultrafiltration membranes, the protein recovery rate can be also improved, microfiltration and ultrafiltration are applied in a combined mode, large molecules and small molecules are respectively removed, protein concentration is also achieved through ultrafiltration, the load of later gel column chromatography separation is reduced, then unwinding after recombinant protein folding is achieved by adding urea, and harmful residues are avoided. The purity of the recombinant human-derived collagen obtained by means of the method can reach up to 99%, the recovery rate is above 65%, the method is simple and convenient to operate, the large-scale production costs of the recombinant human-derived collagen are remarkably reduced, and the method is especially suitable for industrial production.