The cold-shock expression system in Escherichia coli was developed on a manual induction approach using optical density at 600 nm (OD 600 ) measurements and isopropyl β-D-1-thiogalactopyranoside (IPTG) addition. In this study, we show that cold-shock expression performs equally well using an autoinduction approach wherein OD 600 measurements and IPTG addition may be eliminated. We further demonstrate that cold-shock expression with autoinduction can better facilitate high-throughput experiments.