PDF(Pubmed)
Abstract：
UNASSIGNED: Humanization is typically adopted to reduce the immunogenicity of murine antibodies generated by hybridoma technology when used in humans.
UNASSIGNED: Two different strategies of antibody humanization are popularly employed, including \"complementarity determining region (CDR) grafting\" and \"framework (FR) shuffling\" to humanize a murine antibody against human programmed death-1 (PD-1), XM PD1. In CDR-grafting humanization, the CDRs of XM PD-1, were grafted into the human FR regions with high homology to the murine FR counterparts, and back mutations of key residues were performed to retain the antigen-binding affinities. While in FR-shuffling humanization, a combinatorial library of the six murine CDRs in-frame of XM PD-1 was constructed to a pool of human germline FRs for high-throughput screening for the most favorable variants. We evaluated many aspects which were important during antibody development of the molecules obtained by the two methods, including antibody purity, thermal stability, binding efficacy, predicted humanness, and immunogenicity, along with T cell epitope prediction for the humanized antibodies.
UNASSIGNED: While the ideal molecule was not achieved through CDR grafting in this particular instance, FR-shuffling proved successful in identifying a suitable candidate. The study highlights FR-shuffling as an effective complementary approach that potentially increases the success rate of antibody humanization. It is particularly noted for its accessibility to those with a biological rather than a computational background.
UNASSIGNED: The insights from this comparison are intended to assist other researchers in selecting appropriate humanization strategies for drug development, contributing to broader application and understanding in the field.
UNASSIGNED: Two different strategies of antibody humanization are popularly employed, including \"complementarity determining region (CDR) grafting\" and \"framework (FR) shuffling\" to humanize a murine antibody against human programmed death-1 (PD-1), XM PD1. In CDR-grafting humanization, the CDRs of XM PD-1, were grafted into the human FR regions with high homology to the murine FR counterparts, and back mutations of key residues were performed to retain the antigen-binding affinities. While in FR-shuffling humanization, a combinatorial library of the six murine CDRs in-frame of XM PD-1 was constructed to a pool of human germline FRs for high-throughput screening for the most favorable variants. We evaluated many aspects which were important during antibody development of the molecules obtained by the two methods, including antibody purity, thermal stability, binding efficacy, predicted humanness, and immunogenicity, along with T cell epitope prediction for the humanized antibodies.
UNASSIGNED: While the ideal molecule was not achieved through CDR grafting in this particular instance, FR-shuffling proved successful in identifying a suitable candidate. The study highlights FR-shuffling as an effective complementary approach that potentially increases the success rate of antibody humanization. It is particularly noted for its accessibility to those with a biological rather than a computational background.
UNASSIGNED: The insights from this comparison are intended to assist other researchers in selecting appropriate humanization strategies for drug development, contributing to broader application and understanding in the field.
摘要：
■当用于人类时,通常采用人源化来降低由杂交瘤技术产生的鼠抗体的免疫原性。
■普遍采用两种不同的抗体人源化策略，包括“互补决定区（CDR）移植”和“框架（FR）改组”，以人源化针对人类程序性死亡-1（PD-1）的鼠抗体，XMPD1。在CDR移植人源化中，将XMPD-1的CDR移植到与鼠FR对应物高度同源的人FR区，并进行关键残基的回复突变以保留抗原结合亲和力。在FR-改组人性化中，将XMPD-1框内的六个鼠CDR的组合文库构建到人种系FR池中,用于高通量筛选最有利的变体。我们评估了通过两种方法获得的分子的抗体开发过程中重要的许多方面，包括抗体纯度，热稳定性，结合功效，预测人性，和免疫原性，以及人源化抗体的T细胞表位预测。
■虽然在这种特殊情况下，理想的分子不是通过CDR移植实现的，FR-改组在确定合适的候选者方面被证明是成功的。该研究强调FR-改组是一种有效的补充方法，可能会增加抗体人源化的成功率。特别值得注意的是它对那些具有生物学而不是计算背景的人的可访问性。
■此比较的见解旨在帮助其他研究人员为药物开发选择适当的人源化策略，有助于在该领域更广泛的应用和理解。
■普遍采用两种不同的抗体人源化策略，包括“互补决定区（CDR）移植”和“框架（FR）改组”，以人源化针对人类程序性死亡-1（PD-1）的鼠抗体，XMPD1。在CDR移植人源化中，将XMPD-1的CDR移植到与鼠FR对应物高度同源的人FR区，并进行关键残基的回复突变以保留抗原结合亲和力。在FR-改组人性化中，将XMPD-1框内的六个鼠CDR的组合文库构建到人种系FR池中,用于高通量筛选最有利的变体。我们评估了通过两种方法获得的分子的抗体开发过程中重要的许多方面，包括抗体纯度，热稳定性，结合功效，预测人性，和免疫原性，以及人源化抗体的T细胞表位预测。
■虽然在这种特殊情况下，理想的分子不是通过CDR移植实现的，FR-改组在确定合适的候选者方面被证明是成功的。该研究强调FR-改组是一种有效的补充方法，可能会增加抗体人源化的成功率。特别值得注意的是它对那些具有生物学而不是计算背景的人的可访问性。
■此比较的见解旨在帮助其他研究人员为药物开发选择适当的人源化策略，有助于在该领域更广泛的应用和理解。
