Journal article
Purification of prostate-specific membrane antigen using conformational epitope-specific antibody-affinity chromatography
Protein expression and purification, Vol.49(2), pp.251-255
2006
Handle:
https://hdl.handle.net/2376/113612
PMID: 16815035
Abstract
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 Zn
2+ to the equilibration buffer in desalting step provides considerable enhancement in the yield of active PSMA from LNCaP cells.
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Details
- Title
- Purification of prostate-specific membrane antigen using conformational epitope-specific antibody-affinity chromatography
- Creators
- Tiancheng LiuYoko ToriyabeClifford E Berkman
- Publication Details
- Protein expression and purification, Vol.49(2), pp.251-255
- Academic Unit
- Chemistry, Department of
- Publisher
- Elsevier Inc
- Identifiers
- 99900547895601842
- Language
- English
- Resource Type
- Journal article