Khettal B, de Lauzon S, Desfosses B, Ushida S, Marquet A
URA 400, CNRS, UFR Biomédicale des Saints-Pères, Paris, France.
C R Acad Sci III. 1994 May;317(5):381-5.
Antibodies were raised against 3-fluoro-4-aza-estradiol-17-hemisuccinate to elicit abzymes capable of isomerizing delta 5-3-ketosteroids. The hapten was designed to present a planar A ring showing some analogy with the intermediate dienol and a polarity capable of inducing catalytic groups. Antibodies binding the hapten tightly were cloned and purified from either ascites or hybridoma supernatants. Catalytic activity was tested with androst-5-ene-3,17-dione. Among the five different monoclonal antibodies binding the hapten, one proved to enhance significantly the substrate isomerization rate (kcat/kuncat = 830). Initial rates followed Michaelis kinetics and the activity was competitively inhibited by the hapten.
制备了针对3-氟-4-氮杂雌二醇-17-半琥珀酸酯的抗体，以产生能够使δ5-3-酮甾体异构化的抗体酶。该半抗原的设计使其呈现出一个平面A环，与中间二烯醇有一定相似性，并且具有能够诱导催化基团的极性。从腹水或杂交瘤上清液中克隆并纯化了紧密结合该半抗原的抗体。用雄甾-5-烯-3,17-二酮测试催化活性。在结合该半抗原的五种不同单克隆抗体中，有一种被证明能显著提高底物异构化速率（kcat/kuncat = 830）。初始速率遵循米氏动力学，且该活性受到半抗原的竞争性抑制。
