Tawfik D S, Green B S, Chap R, Sela M, Eshhar Z
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Proc Natl Acad Sci U S A. 1993 Jan 15;90(2):373-7. doi: 10.1073/pnas.90.2.373.
The low abundance and activity of catalytic antibodies are major obstacles to their selection from the virtually unlimited repertoire of antibody binding sites. The requirement for new screening methodologies is further emphasized by the availability of combinatorial libraries, in which a functional polypeptide has to be selected out of millions of possibilities. We present a simple and sensitive screening approach (termed catELISA) based on immobilized substrates and immunodetection of the end product of the catalyzed reaction. The feasibility of catELISA is demonstrated here by the generation of potent ester-hydrolyzing antibodies by direct screening of hybridoma supernatants. We show that this approach is not only facile but general: it is not limited by type of reaction, substrate, or catalyst (enzymes, catalytic antibodies, chemical catalysts). catELISA opens a route to catalytic antibodies that replaces existing lengthy and arduous methods, thus allowing us to expand their number and improve their quality and to address questions that would otherwise be difficult to answer.
催化抗体的低丰度和活性是从几乎无限的抗体结合位点库中筛选它们的主要障碍。组合文库的出现进一步凸显了对新筛选方法的需求，在组合文库中，必须从数百万种可能性中选出一种功能性多肽。我们提出了一种基于固定化底物和对催化反应终产物进行免疫检测的简单且灵敏的筛选方法（称为催化酶联免疫吸附测定，catELISA）。通过直接筛选杂交瘤上清液产生高效酯水解抗体，证明了catELISA的可行性。我们表明，这种方法不仅简便而且通用：它不受反应类型、底物或催化剂（酶、催化抗体、化学催化剂）的限制。catELISA为催化抗体开辟了一条途径，取代了现有的冗长而艰巨的方法，从而使我们能够增加其数量、提高其质量，并解决那些原本难以回答的问题。
