کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2021949 1069271 2006 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Purification of prostate-specific membrane antigen using conformational epitope-specific antibody-affinity chromatography
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Purification of prostate-specific membrane antigen using conformational epitope-specific antibody-affinity chromatography
چکیده انگلیسی

Prostate-specific membrane antigen (PSMA) is a type II membrane protein that has attracted significant attention as a target for immunioscintigraphic and radioimmunotherapeutic applications for prostate cancer. However, definitive studies on its substrate and inhibitor specificity as well as protein–protein interactions have been somewhat limited by difficulties in the purification of native PSMA. In this study, we optimized the purification of native PSMA from LNCaP cells using conformational epitope-specific antibody-affinity chromatography. Western blot analysis and an HPLC-based enzymatic activity assay were used to compare the yield and activity of PSMA purified by different methods. The ratio of purified PSMA in a native and active conformation was determined by quantifying the amount of non-native PSMA not retained in a second antibody-affinity isolation. The addition of both a neutralization step and the inclusion of Zn2+ to the equilibration buffer in desalting step provides considerable enhancement in the yield of active PSMA from LNCaP cells.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Protein Expression and Purification - Volume 49, Issue 2, October 2006, Pages 251–255
نویسندگان
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