利用重链互补决定区 3 移植结合体外体细胞超突变实现抗体的人源化。
Humanization of antibodies using heavy chain complementarity-determining region 3 grafting coupled with in vitro somatic hypermutation.
机构信息
Anaptysbio Inc., San Diego, California 92131.
Anaptysbio Inc., San Diego, California 92131.
出版信息
J Biol Chem. 2013 Mar 15;288(11):7688-7696. doi: 10.1074/jbc.M112.445502. Epub 2013 Jan 25.
Abstract
A method for simultaneous humanization and affinity maturation of monoclonal antibodies has been developed using heavy chain complementarity-determining region (CDR) 3 grafting combined with somatic hypermutation in vitro. To minimize the amount of murine antibody-derived antibody sequence used during humanization, only the CDR3 region from a murine antibody that recognizes the cytokine hβNGF was grafted into a nonhomologous human germ line V region. The resulting CDR3-grafted HC was paired with a CDR-grafted light chain, displayed on the surface of HEK293 cells, and matured using in vitro somatic hypermutation. A high affinity humanized antibody was derived that was considerably more potent than the parental antibody, possessed a low pm dissociation constant, and demonstrated potent inhibition of hβNGF activity in vitro. The resulting antibody contained half the heavy chain murine donor sequence compared with the same antibody humanized using traditional methods.
摘要
一种同时对单克隆抗体进行人源化和亲和力成熟的方法已经开发出来，该方法使用重链互补决定区（CDR）3 移植结合体外体细胞高频突变。为了在人源化过程中最小化使用的鼠源性抗体序列的量，仅将识别细胞因子 hβNGF 的鼠抗体的 CDR3 区域移植到非同源的人胚系 V 区。所得的 CDR3 移植 HC 与 CDR 移植的轻链配对，展示在 HEK293 细胞表面，并通过体外体细胞高频突变进行成熟。衍生出一种高亲和力的人源化抗体，其比亲本抗体具有更高的效力，具有低 pm 解离常数，并在体外显示出对 hβNGF 活性的有效抑制。与使用传统方法人源化的相同抗体相比，所得抗体的重链鼠供体序列减少了一半。
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