HIV 1 mutation results in immune evasion, which presents a serious challenge for conventional strategies for developing effective vaccines. So far, much experimental evidence indicates that HIV 1 particles in the blood of patients can be cleaned principally by neutralizing antibodies. Based on these facts, we prepared triple combination of epitope vaccines with the objective of inducing antibodies with predefined multi epitope specificity against HIV 1. According to the sequences of three neutralizing epitopes (RILAVERYLKD, ELDKWA and GPGRAFY, designated E1, E2, and E3, respectively) on HIV 1 envelope proteins, three epitope peptides ((E1)2: C (RILAVERYLKDG) 2; (E2)4: C (ELDKWAG) 4; and (E3)2: C (GPGRAFY) 2) were synthesized and then conjugated with carrier protein keyhole limpet hemocyanin (KLH) or bovine serum albumin (BSA), and used for immunizing rabbits. After the vaccine course, the triple combination of epitope vaccines induced high levels of predefined multi epitope specific antibodies. An immunoblotting analysis demonstrated that the antibodies could recognize the native epitopes on both gp41 protein and V3 loop peptide. Furthermore, we compared the immune responses of three doses of epitope peptides in the candidate epitope vaccine. Strong antibody responses to three epitopes were observed in a dose dependent manner, with increasing dose raising the immune response. This result indicated that immunotolerance did not occur using an epitope vaccine dose of 80 μg. Thus, our results demonstrate that epitope vaccines in combination can synchronously induce high levels of antibodies with predefined multi epitope specificity against HIV 1, and may be used to develop effective vaccines against HIV as a new strategy. HIV 1 mutation results in immune evasion, which presents a serious challenge for conventional strategies for developing effective vaccines. So far, much experimental evidence that that HIV 1 particles in the blood of patients can be cleaned principally by neutralizing antibodies. Based on these facts, we prepared triple combination of epitope vaccines with the objective of inducing antibodies with predefined multi-specificity specificity against HIV 1. According to the sequences of three neutralizing epitopes (RILAVERYLKD, ELDKWA and GPGRAFY, designated E1, E2, and E3, respectively) on HIV 1 envelope proteins, three epitope peptides ((E1) 2: C (RILAVERYLKDG) 2; (E2) 4: C (ELDKWAG) 4; and (E3) 2: C (GPGRAFY) 2) were synthesized and then conjugated with carrier protein keyhole limpet hemocyanin (KLH) or bovine serum albumin (BSA), and used for immunizing rabbits. After the vaccine course, the triple combination of epitope vaccines induced high levels of predefined multi epitope specific antibodies. An immunoblotting analysis demonstrated that the antibodies could recognize the native epitopes on both gp41 protein and V3 loop peptide. Further, we compared the immune responses of three doses of epitope peptides in the candidate epitope vaccine. This result indicates that immunotolerance did not occur using antigen vaccine dose of 80 μg. Thus, our results demonstrate that epitope vaccines in combination can synchronously induce high levels of antibodies with predefined multi specificity specificity against HIV 1, and may be used to develop effective vaccines against HIV as a new strategy.