The marine algal biomass of Ulva lactuca was utilized for the extraction of cellulose and the development of cellulose nanocrystals/graphene oxide film. Cellulose nanocrystals with 50–150 nm were produced by H2SO4 hydrolysis of the algal cellulose. The adsorption efficiency of the nanocomposite film for Fe(II) and Fe(III) ions was successfully evaluated using Box-Behnken design. The maximum removal for Fe(II) (64.15%) could be attained at pH 5.13, adsorbent dosage 7.93 g L−1 and Fe(II) concentration 15.39 mg L−1, while the biosorption of Fe(III) was 69.92% at pH 5.0, adsorbent dosage 2 g L−1, and Fe(III) concentration 15.0 mg L−1. However, in the binary system, the removal efficiency of Fe(II) was enhanced to 95.48% at Fe(II):Fe(III) ratio of 1:1, while the Fe(III) removal was increased to 79.17% at ratio 1:2. The pseudo-second-order kinetics exhibited better fitting to the experimental results of Fe(II) and Fe(III) adsorption in both single and binary systems. The intra-particle diffusion was prominent during the biosorption, but the effect of the external mass transfer was significant. The Langmuir, Freundlich, Langmuir–Freundlich, Temkin, and Dubinin-Radushkevich isotherms showed satisfactory fitting to the experimental data, but they differ in priority based on iron state and pH. The adsorption of Fe(II) in the presence of Fe(III) in a mixture was best represented by the extended Langmuir model, while the extended Langmuir–Freundlich model best fitted the adsorption of Fe(III). The FT-IR analysis indicated that physisorption through electrostatic interaction/complexation is the predominant mechanism for the adsorption of iron using the nanocomposite film.