Generation of single domain antibody fragments derived from camelids and generation of manifold constructs.
Generation of single domain antibody fragments derived from camelids and generation of manifold constructs.
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DOI：
10.1007/978-1-61779-974-7_8
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发表时间：
2012-01-01
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影响因子：
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通讯作者：
Muyldermans, Serge
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文献类型：
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作者：
Vincke, Cecile;Gutierrez, Carlos;Muyldermans, Serge
用可溶性、正确折叠的蛋白质免疫骆驼科动物（骆驼和美洲驼），在独特的骆驼科动物仅重链抗体（HCAb）中产生亲和力成熟的免疫应答。免疫动物的外周血淋巴细胞用于从噬菌体展示载体中的HCAb克隆抗原结合抗体片段。这些抗原结合片段的文库的代表性等分试样用于通过连续轮淘选来检索单结构域抗原特异性结合剂。串联克隆这些单结构域抗体片段以产生多种构建体（二价、双互补位或双特异性构建体）以增加其功能亲和力、增加特异性或连接两个独立的抗原分子。
Immunizing a camelid (camels and llamas) with soluble, properly folded proteins raises an affinity-matured immune response in the unique camelid heavy-chain only antibodies (HCAbs). The peripheral blood lymphocytes of the immunized animal are used to clone the antigen-binding antibody fragment from the HCAbs in a phage display vector. A representative aliquot of the library of these antigen-binding fragments is used to retrieve single domain antigen-specific binders by successive rounds of panning. These single domain antibody fragments are cloned in tandem to generate manifold constructs (bivalent, biparatopic or bispecific constructs) to increase their functional affinity, to increase specificity, or to connect two independent antigen molecules.