Chitosan, as a potential vaccine delivery material, has obtained much attention for immunization prevention and therapy. However, its poor water solubility brings inconvenience for the practical applications. To address this issue, researchers have carried out many chemical modifications to prepare water-soluble chitosan derivatives for vaccine delivery. In this work, we prepared a chitosan derivative N-phosphonium chitosan with excellent water solubility and explored its potential as an intramuscular vaccine delivery system by using ovalbumin as a model antigen. Different vaccine formulations were intramuscularly injected into test mice. Through an immunohistochemistry assay, N-phosphonium chitosan-based antigen formulation could promote antigen arrival from injection site to the secondary lymph organ spleen. Further immunization results showed that 1 mg/ml N-phosphonium chitosan-based vaccine formulation could contribute to significantly higher level of antigen-specific immune responses, including higher antigen-specific IgG antibody titer, splenocyte proliferation, and cytokines secretion (interferon-γ, interleukin-10, and interleukin-4) by the splenocytes of the immunized mice. From the results, the water-soluble chitosan derivative N-phosphonium chitosan could be developed as a potential antigen carrier for immunization prevention and therapy.