Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama
Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama
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DOI：
10.1016/s0022-1759(00)00188-5
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发表时间：
2000-06-23
影响因子：
2.2
通讯作者：
Frenken, L
中科院分区：
文献类型：
--
作者：
van der Linden, R;de Geus, B;Frenken, L
到目前为止，功能性重链免疫球蛋白只在骆驼和大羊驼身上发现。这些重链igg （V-HH）的抗原特异性片段在生物技术领域引起了极大的兴趣，因为它们非常稳定，可以由酵母（Saccharomyces cerevisiae）高水平生产。本文描述的工作是为了确定美洲驼（Lama glama）是否是抗原特异性V-HH片段的实际来源。用不同抗原对大羊驼进行免疫，并在免疫过程中检测抗体反应。常规和重链IgG抗体对每一种抗原都产生反应。重链IgG库显示出与传统羊驼igg不同的识别模式，导致可达表位库的扩增。大羊驼血清中重链IgG抗体的比例低于骆驼。为了从外周血b细胞中特异性、高效地分离出V-HH基因，我们测定了Lama glama重链IgG的长、短铰链序列，揭示了一种新的短铰链重链IgG亚类的存在。设计了长、短铰链特异性PCR引物，用于羊驼V-HH文库的构建。我们的结论是，使用所描述的技术，抗原特异性V-HH抗体片段很容易从羊驼中获得，从而为各种应用提供了非常有价值的结合分子。(C) 2000 Elsevier Science B.V.版权所有
Functional heavy chain immunoglobulins have, so far, only been found in camels and llamas. Antigen-specific fragments of these heavy chain IgGs (V-HH) are of great interest in biotechnology because they are very stable and can be produced at high level by the yeast Saccharomyces cerevisiae. The work described in this paper was conducted to determine whether llamas (Lama glama) are a practical source of antigen-specific V-HH fragments. Llamas were immunised with various types of antigens and the antibody responses were examined during the course of immunisation. Both, conventional and heavy chain IgG antibodies were produced in response to each of the antigens. The heavy chain IgG repertoire displayed a recognition pattern different to that of conventional llama IgGs, resulting in the expansion of the accessible epitope repertoire. Llamas have a lower proportion of heavy chain IgG antibodies in their serum than have camels. To enable the specific and efficient isolation of V-HH genes from peripheral blood B-cells, the long and short-hinge sequences of Lama glama heavy chain IgGs were determined, revealing the presence of a novel subclass of short-hinge heavy chain IgG. Long and short-hinge specific PCR primers were designed to be used in the construction of llama V-HH libraries. We conclude that, using the techniques described, antigen-specific V-HH antibody fragments are readily accessible from the llama, thus providing highly valuable binding molecules for a variety of applications. (C) 2000 Elsevier Science B.V. All rights reserved.