Monoclonal antibodies，one of the most widely used tools in biological research, with the potential soon to become important therapeutics, have been with us for just over 40 years. These antibodies are monovalent, bind to a single epitope and are produced by a single B-lymphocyte clone. The first monoclonal antibody was generated in mice in 1975 by Milstein and Kohler, using the hybridoma technique. 

单克隆抗体是生物学研究中使用较广泛的工具之一，也具有成为重要治疗药物的潜力，它的出现已有 40 多年的历史。这些抗体是单价的，与单个表位结合并且由单个 B 淋巴细胞克隆产生。1975 年，Milstein 和 Kohler 使用杂交瘤技术在小鼠中产生了第一个单克隆抗体。

1.History and production 历史和产生

Milstein and Kohler realised that in order to get an antibody where each molecule was identical to one another, they could harvest single B-cells from the spleen of a mouse and fuse them to myeloma (cancer) cell line, producing hybridomas.Antibody-producing B-cells are clonal, derived from memory B-cells, which means that every B-cell generated from a single memory B-cell produces the exact same antibody. These are termed ‘Monoclonal antibodies’ (single clone).

Milstein 和 Kohler 意识到，为了获得每个分子彼此相同的抗体，他们从小鼠脾脏中收获单个 B 细胞并将其融合到骨髓瘤（癌症）细胞系中，从而产生杂交瘤。产生抗体的 B 细胞是来自记忆 B 细胞的克隆，这意味着从单个记忆 B 细胞产生的每个 B 细胞产生相同的抗体。这些被称为「单克隆抗体」（单克隆）。

Hybridoma production is still the most common method of Monoclonal antibody production. The fusions that are stable enough to grow in vitro are selected, sub-cloned and form a monoclonal-antibody producing hybridoma. These clones are then cultured and the antibodies they produce are purified either from the cells themselves, or the cell culture lysates. Each monoclonal antibody is defined by its unique binding site on the target protein, and this unique binding determines its specificity.

杂交瘤生产仍然是单克隆抗体生产中常用的方法。选择足够稳定以在体外生长的融合体，亚克隆并形成产生单克隆抗体的杂交瘤。然后培养这些克隆，并将它们产生的抗体从细胞本身或细胞培养裂解物中纯化。每种单克隆抗体由其在靶蛋白上的结合位点定义，并且这种的结合决定了其特异性。

 

2.Uses and alternative production methods 用途和替代生产方法

Antibodies with this level of specificity are a popular research tool for the detection of a single protein. They can also be used as a probe to locate the protein of interest inside a cell or organ. In recent years, alternative methods of producing monoclonal antibodies have become available. The so-called phage-display method meant that recombinant versions of the antigen-binding part of the antibody could be screened for specificity from a library of fusions to a carrier phage protein. Following on, further refinements to recombinant antibody production resulted in cell line-producing recombinant antibodies where the species of the constant region can be customized. The advantage of this new production method is that the issues around long-term stability of a hybridoma, with potential loss of a precious clone, is avoided.

The most important reason to use a monoclonal antibody has to be its consistency. As the cell-line is essentially immortal and unchanging, no new animals are needed for further production and the antibody remains consistent from batch to batch in its performance.

具有这种特异性水平的单克隆抗体是检测单一蛋白质中非常流行的研究工具。它们也可以用作探针来定位细胞或器官内感兴趣的蛋白质。近年来，生产单克隆抗体的替代方法也已经产生。如所谓的噬菌体展示方法，即从融合文库筛选特异性抗体的抗原结合部分，以重组形式与噬菌体蛋白质进行融合表达。接下来，对重组抗体生产的进一步改进而产生细胞系的重组抗体，其中恒定区的种类可以定制。这种新的生产方法的优点是克服了杂交瘤长期不稳定性可能会损失宝贵的克隆的问题。

使用单克隆抗体最重要的原因是它的一致性。由于细胞系基本上是不朽且不变的，所以不需要新的动物用于进一步生产，并且抗体在其性能中从批次到批次保持一致。
 

In addition to the use of the entire immunoglobulin, for certain experiments these immunoglobulin molecules can be cleaved to give smaller versions with specific properties. The the constant region (Fc) of the immunoglobulin can be cleaved either above or below the hinge region, leaving either (Fab’)2 or Fab fragments. Since the Fc portion has affinity to Ig receptors, the removal of this part of the antibody reduces non-specific binding and is therefore often used in Flow cytometry applications or where binding to Fc receptors must be avoided.

除了使用完整的免疫球蛋白之外，对于某些实验，这些免疫球蛋白分子可以被切割以产生具有特定性质的片段。免疫球蛋白的恒定区（Fc）可以在铰链区上方或下方切割，留下（Fab'）2 或 Fab 片段。由于 Fc 部分对 Ig 受体具有亲和力，所以抗体这部分的去除减少了非特异性结合，因此经常这种抗体常用于流式细胞术应用或者其他必须避免结合 Fc 受体的免疫实验。

Biorbyt 拥有的人，小鼠，大鼠，兔各种来源的单克隆抗体，优势和克隆统计数据如下。感兴趣的请联系我们，获取更多单克隆抗体的产品信息和咨询。
 

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