Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs)
Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs)
复制标题
DOI：
10.1016/j.jim.2007.04.008
复制
发表时间：
2007-07-31
影响因子：
2.2
通讯作者：
Shoemaker, Charles B.
中科院分区：
文献类型：
--
作者：
Maass, David R.;Sepulveda, Jorge;Shoemaker, Charles B.
与从常规IgG制备的单链抗体相比，来源于不寻常的骆驼科动物仅重链IgG类（HCAbs）的重组单结构域抗体片段（VHH）具有许多有利的性质。因此，VHH已被广泛用作结合试剂，并开始显示出作为治疗剂的潜力。迄今为止，VHH遗传物质的来源一直是骆驼和美洲驼，尽管它们体型庞大且可用性有限。在这里，我们证明，更小，更听话和广泛使用的羊驼是一个很好的来源VHH编码DNA。羊驼血清IgG由约50%的HCAb组成，主要是短铰链品种。对编码超过50个随机VHH和铰链结构域的DNA进行测序允许设计PCR引物，所述PCR引物将扩增几乎所有羊驼VHH编码DNA用于噬菌体展示文库构建。用绵羊肿瘤坏死因子α（TNF α）免疫羊驼，从排出免疫部位的淋巴结制备VHH噬菌体展示文库，并成功地用于分离结合和中和绵羊TNF α的VHH。(C)2007 Elsevier B.V.保留所有权利。
Recombinant single domain antibody fragments (VHHs) that derive from the unusual camelid heavy chain only IgG class (HCAbs) have many favourable properties compared with single-chain antibodies prepared from conventional IgG. As a result, VHHs have become widely used as binding reagents and are beginning to show potential as therapeutic agents. To date, the source of VHH genetic material has been camels and llamas despite their large size and limited availability. Here we demonstrate that the smaller, more tractable and widely available alpaca is an excellent source of VHH coding DNA. Alpaca sera IgG consists of about 50% HCAbs, mostly of the short-hinge variety. Sequencing of DNA encoding more than 50 random VHH and hinge domains permitted the design of PCR primers that will amplify virtually all alpaca VHH coding DNAs for phage display library construction. Alpacas were immunized with ovine tumour necrosis factor a (TNF alpha) and a VHH phage display library was prepared from a lymph node that drains the sites of immunizations and successfully employed in the isolation of VHHs that bind and neutralize ovine TNF alpha. (C) 2007 Elsevier B.V. All rights reserved.