Eliciting antibodies that neutralize a broad range of circulating HIV strains (broadly neutralizing antibodies bnAbs) represents a key priority for vaccine development. HIV superinfection (re-infection with a second strain following an established infection) has been associated with neutralization breadth, and can provide insights into how the immune system responds to sequential exposure to distinct HIV envelope glycoproteins (Env). Characterizing the neutralizing antibody (nAb) responses in four superinfected women revealed that superinfection does not boost memory nAb responses primed by the first infection or promote nAb responses to epitopes conserved in both infecting viruses. While one superinfected individual developed potent bnAbs, superinfection was likely not the driver as the nAb response did not target an epitope conserved in both viruses. Rather, sequential exposure led to nAbs specific to each Env but did not promote bnAb development. Thus, sequential immunization with heterologous Envs may not be sufficient to focus the immune response onto conserved epitopes. Display omitted •HIV superinfection does not efficiently recruit cross-reactive memory B cells•Superinfection results in antibody responses specific to each infecting strain•No evidence that superinfection drives the development of bnAbs HIV-infected individuals can be re-infected with a second strain (superinfection), providing a model that informs the use of sequential immunizations in future vaccines. Sheward et al. find that superinfection fails to boost memory B cells primed by the first infection and does not promote broadly neutralizing antibodies to HIV.