POLYCLONAL LIBRARY--RESOURCES FOR ANTIBODIES & CATALYSIS

对特定反应具有催化活性的抗体可以是 在Balb/c小鼠免疫后，用适当的 过渡态模拟。然而，无法准确地确定 许多反应的状态是一般探索的障碍。 抗体催化的新现象。绕过将军的一条路 缺乏关于过渡态的准确结构信息可能 通过采用另一种方法搜索大样本来找到 Balb/c的抗体谱系可以催化特定的 反应。 这项提议旨在展示大型企业的多功能性和力量 杂交瘤文库整合了很大一部分 Balb/c小鼠的特异性(非克隆型)谱系。这个 图书馆，其组成基本上正在进行中，由 聚乙二醇化多克隆活化B细胞融合的研究 未免疫的Balb/c小鼠。该图书馆最终将包含一个大型 离散杂交瘤种群数量(约10.000)，每个杂交瘤种群 由至少1个克隆组成，通常由3到5个或更多克隆组成。这个图书馆 将被筛选以证明它的一些成员显示出催化作用 活动。将使用显色或放射性底物来识别 杂交瘤种群催化任何一种 多种多样的结构主题。此外，还有一种可能性是 在其成员中，人们可以找到不同的抗体谱，其 亲和力或抗原特异性使它们具有实用价值。多样性 在文库中所代表的抗原反应性的将由 针对抗原小组筛选其成员，该抗原小组包括 分化抗原、不同细菌种类和解决方案 可溶性抗原(甲状腺素、胰岛素、钙调素和地高辛) 亚微摩尔浓度。这种方法的意义在于 它的潜力提供了一种替代途径，以推导出 免疫程序的抗体。此外，还有耐人寻味的 正常情况下表达被阻断的抗体可能 这项技术可以获得对容差的限制。这 可能会产生高度保守的抗体 特定类型的分子或特定类别的分子的决定因素 对许多不同物种感兴趣的细胞。

Antibodies with catalytic activity for a particular reaction can be elicited in the Balb/c mouse after immunization with an appropriate transition state analog. However, the inability to determine precisely the states of many reactions is a barrier to the general exploration of the novel phenomenon of antibody catalysis. A way around the general lack of precise structural information about the transition state might be found by pursuing the alternative approach of searching a large sample of the Balb/c repertoire for antibodies which can catalyze a particular reaction. This proposal seeks to demonstrate the versatility and power of a large hybridoma library which incorporates a significant fraction of the repertoire of specificities (not clonotypes) in the Balb/c mouse. The library, whose composition is substantially underway, is constructed by the PEG-assisted fusion of polyclonally activated B cell blasts from unimmunized Balb/c mice. The library will ultimately contain a large number (ca. 10.000) of discrete hybridoma populations, each of which is composed of at least 1 and usually 3 to 5 or more clones. This library will be screened to demonstrate that some of its members show catalytic activity. Chromogenic or radioactive substrates will be used to identify hybridoma populations that catalyze the hydrolysis of any one of a variety of structural motifs. In addition there is the possibility that among its members one can find a diverse spectrum of antibodies whose affinity or antigen specificity qualifies them as useful. The diversity of antigen reactivities represented in the library will be determined by screening its members against an antigen panel that includes differentiation antigens, different bacterial species, and solutions of soluble antigens (thyroxine, insulin, calmodulin, and digoxin) at submicromolar concentrations. The significance of this approach lies in its potential to provide an alternative route to the derivation of antibodies to immunization programs. Also, there is the intriguing possibility that antibodies whose expression is normally blocked by constraints of tolerance could be accessed by this technology. This might make it possible to derive antibodies to highly conserved determinants on particular types of molecules or specific categories of cells that are of interest in many different species.