论文部分内容阅读
Objective To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9/recombinant human interleukin 2 (rhIL 2) conjugate Methods MoAb 520C9 reactive with the protooncogene c erbB 2 product P185 was chemically conjugated with rhIL 2 by using a simple two step method First, the rhIL 2 was activated by Sulfosuccinimidyl 4 [N maleimidomethyl] cyclohexane 1 carboxylate, a heterobifunctional linker, and N succinimidyl s acetylthioacetate was introduced onto 520C9 Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL 2 to generate 520C9 rhIL 2 immunoconjugate Results The immunoconjugate retained the antigen binding activity compared to the respective native antibody as determined by an indirect live cell binding assay The immunoconjugate also possessed IL 2 activity as measured by the standard CTLL 2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine activated killer cells Conclusion Our method of conjugation of rhIL 2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL 2 This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines
Objective To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9 / recombinant human interleukin 2 (rhIL 2) conjugate Methods MoAb 520C9 reactive with the protooncogene c erbB 2 product P185 was chemically conjugated with rhIL 2 by using a simple two step method First, the rhIL 2 was activated by Sulfosuccinimidyl 4 [N maleimidomethyl] cyclohexane 1 carboxylate, a heterobifunctional linker, and N succinimidyl s acetylthioacetate was introduced onto 520C9 Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL 2 to generate 520C9 rhIL 2 immunoconjugate Results The immunoconjugate retained the antigen binding activity compared to the corresponding native cell as determined by an indirect live cell binding assay The immunoconjugate also possessed IL 2 activity as measured by the standard CTLL 2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine ac tivated killer cells Conclusion Our method of conjugation of rhIL 2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL 2 This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines