SummaryPrawn muscles were treated with acetic acid and high‐pressure processing (600 MPa) separately to analyse their antigenicity and immunogenicity. The protein fractions were separated and isolated using preparative HPLC, and their antigenicity was analysed using Immunoglobulin G (IgG) ELISA kit. Out of thirty‐nine protein fractions, only four (A10, A11, B10 and C9) were detected with antigenic potentials. The immunogenicity of these protein fractions was analysed using human PBMCs, and supernatants were collected at multiple times from 0 to 144 h. The treated fractions (B10 and C9) analysed using Immunoglobulin E (IgE) ELISA kit showed significantly (P < 0.05) lower pro‐ and anti‐inflammatory cytokine production compared with control (A10). The allergenic fractions were characterised using an LC/MS/MS, which identified nine proteins. Among these, six proteins (tropomyosin, arginine kinase, haemocyanin, enolase, vitellogenin and 14‐3‐3 zeta) have been established as allergenic in prawn muscle and ovaries. Other three proteins (beta‐1,3‐glucan‐binding protein, translationally controlled tumour protein and farnesoic acid O‐methyltransferase short isoform protein) identified in this study need further investigation for their immunogenic properties.