کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2833471 1163873 2007 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A phage antibody to the active site of human placental alkaline phosphatase with higher affinity to the enzyme–substrate complex
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شناسی مولکولی
پیش نمایش صفحه اول مقاله
A phage antibody to the active site of human placental alkaline phosphatase with higher affinity to the enzyme–substrate complex
چکیده انگلیسی

Selection of specific antibodies from large repertoires is of importance in generating antibodies to specific structural determinants and in studying structure–function relationships. Alkaline phosphatase (AP) has several isozymes with various degrees of homology and a range of common synthetic substrates. We have previously reported the generation of isozyme specific anti-enzyme antibodies to an oncofetal antigen, placental alkaline phosphatase (PLAP) by using a specific uncompetitive inhibitor, l-Phe-Gly-Gly along with the substrate para-nitrophenyl phosphate (pNPP), to elute scFvs from a phage-displayed immunoglobulin library. These antibodies were directed to the active site and inhibited enzyme activity. An uncompetitive inhibitor acts by stabilizing the enzyme–substrate (ES) complex. In the present work, we report the characteristics of a clone VE5, selected by the same method. This clone has a higher binding affinity for ES complex than for enzyme alone. This is true for all the three isozymes (placental, bone and intestinal) tested. However, the other synthetic small molecular substrate, disodium phenyl phosphate inhibits phage binding. The clone possibly binds to the conserved structures of the active site of the AP isozymes and the higher affinity binding to AP–pNPP complex reflects the method of selection. Such anti-enzyme antibodies have a possible potential role in dissecting structure–function relationship of enzymatic antigens.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Molecular Immunology - Volume 44, Issue 4, January 2007, Pages 369–376
نویسندگان
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