Antibodies Engineered for Allosteric Control of Binding.

描述（由申请人提供）：最新的纯化方法 由于需要多个步骤和经验， 发展原则上，免疫亲和层析（IAC）应提供 一个非常有吸引力的选择，因为它提供了显着丰富的单一 步骤，并容易地适应任何目标化合物。不幸的是，IAC 由于难以洗脱捕获的配体， 而不使抗体和/或产物失活。一般来说，大量 必须筛选候选抗体，以找到释放抗原的抗体 在足够温和的条件下。即使这样，洗脱通常会导致一些 抗体失活，使得IAC操作昂贵，因为需要 定期更换列。这项拟议中的研究将测试 能够对任何scFv施加变构调节的分子设计。洗脱 利用该技术的IAC的条件是真正温和的变构 调节剂结合导致scFv采用低活性构象， 通过其专门结构所固有的灵活性，可以随时获得。 因此，不发生scFv解折叠，并且应保留全部活性 经过多次色谱循环。此外，设计的有效性是 预期有助于利用甚至非常高亲和力的scFv（例如，10-11 M）在IAC中。 拟议商业应用：不可用

DESCRIPTION (provided by applicant): Most current purification methodologies are very expensive due to the requirement for multiple steps and empirical development. In principle, immuno-affinity chromatography (IAC) should provide a very attractive alternative as it affords remarkable enrichment in a single step and is easily adapted to any target compound. Unfortunately however, IAC has seen rather restricted use due to difficulty eluting captured ligand without inactivating the antibody and/or product. Generally a large number of candidate antibodies must be screened to find one that releases the antigen under sufficiently mild conditions. Even then, elution usually causes some antibody inactivation, making IAC operationally expensive due to the need for periodic column replacement. The proposed research will test efficacy of a molecular design capable of imposing allosteric regulation on any scFv. Elution conditions of IAC utilizing this technology are truly mild as allosteric regulator binding causes the scFv to adopt a low activity conformation that is readily accessible through flexibility inherent in its specialized structure. Consequently, no scFv unfolding occurs and full activity should be retained over numerous chromatography cycles. Furthermore, efficacy of the design is expected to facilitate utilization of even very high affinity scFv (eg. 10-11 M) in IAC. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE