Water alternating gas (WAG) flooding is a widely employed enhanced oil recovery method in various reservoirs worldwide. In this research, we will employ SiO2 nanofluid alternating with the CO2 injection method as a replacement for the conventional WAG process in oil flooding experiments. The conventional WAG method suffers from limitations in certain industrial applications, such as extended cycle times, susceptibility to water condensation and agglomeration, and ineffectiveness in low-permeability oil reservoirs, thus impeding the oil recovery factor. In order to solve these problems, this study introduces SiO2 nanofluid as a substitute medium and proposes a SiO2 nanofluid alternate CO2 flooding method to enhance oil recovery. Through the microcharacterization of SiO2 nanofluids, comprehensive evaluations of particle size, dispersibility, and emulsification performance were conducted. The experimental results revealed that both SiO2-I and SiO2-II nanoparticles exhibited uniform spherical morphology, with particle sizes measuring 10−20 nm and 50−60 nm, respectively. The SiO2 nanofluid formulations demonstrated excellent stability and emulsification properties, highlighting their potential utility in petroleum-related applications. Compared with other conventional oil flooding methods, the nanofluid alternating CO2 flooding effect is better, and the oil flooding effect of smaller nanoparticles is the best. Nanofluids exhibit wetting modification effects on sandstone surfaces, transforming their surface wettability from oil-wet to water-wet. This alteration reduces adhesion forces and enhances oil mobility, thereby facilitating improved fluid flow in the rock matrix. In the oil flooding experiments with different slug sizes, smaller gas and water slug sizes can delay the breakthrough time of nanofluids and CO2, thereby enhancing the effectiveness of nanofluid alternate CO2 flooding for EOR. Among them, a slug size of 0.1 PV approaches optimal performance, and further reducing the slug size has limited impact on improving the development efficiency. In oil flooding experiments with different slug ratios, the optimal slug ratio is found to be 1:1. Additionally, in oil flooding experiments using rock cores with varying permeability, lower permeability rock cores demonstrate higher oil recovery rates.