Kinetic exclusion assay technology: Characterization of molecular interactions
Kinetic exclusion assay technology: Characterization of molecular interactions
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DOI：
10.1089/adt.2004.2.647
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发表时间：
2004-12-01
影响因子：
1.8
通讯作者：
Brault, PA
中科院分区：
文献类型：
--
作者：
Darling, RJ;Brault, PA
在亲和力、结合动力学、化学计量学、特异性和热力学方面表征分子间相互作用可以促进在蛋白质治疗剂的发现和开发中选择先导化合物。（Sapidyne Instruments，Inc.，博伊西，ID）是一种相对较新的技术，其在表征分子相互作用，特别是关于抗体治疗剂方面获得使用。KinExA提供了一个平台，允许在溶液相中使用未修饰的分子测量真正的平衡结合亲和力和动力学。这是通过使用固相固定化分子在允许足够的时间达到平衡后（亲和力测量）或在预平衡条件下（动力学）探测一种相互作用组分的自由浓度来实现的。在这篇综述中，KinExA技术背后的理论进行了讨论，并提供了应用该技术的抗体表征的例子。最后，KinExA，Biacore（表面等离子体共振），和等温滴定量热法之间的比较，并讨论了KinExA的潜在的未来改进和应用。
Characterization of intermolecular interactions in terms of affinity, binding kinetics, stoichiometry, specificity, and thermodynamics can facilitate the selection of lead compounds in the discovery and development of protein therapeutics. KinExA(R) ( Sapidyne Instruments, Inc., Boise, ID) is a relatively new technology that is gaining use in characterizing molecular interactions, particularly with respect to antibody therapeutics. KinExA offers a platform that allows the measurement of true equilibrium binding affinity and kinetics using unmodified molecules in solution phase. This is accomplished by using a solid- phase immobilized molecule to probe for free concentration of one interaction component after allowing sufficient time to reach equilibrium ( affinity measurements), or under pre- equilibrium conditions ( kinetics). In this review, the theory behind KinExA technology is discussed, and examples of applying this technology to antibody characterization are provided. Finally, a comparison among KinExA, Biacore ( surface plasmon resonance), and isothermal titration calorimetry is presented, and potential future improvements and applications of KinExA are discussed.