Immunopeptidometric assay for enzymatically active prostate-specific antigen.

Abstract:

BACKGROUND:Determinations of certain forms of prostate-specific antigen (PSA) have been shown to increase the specificity for prostate cancer (PCa). One such variant, proteolytically active PSA, is a potentially useful tumor marker, but it is not specifically recognized by antibodies. Using phage display libraries, we previously identified a "family" of peptides that bind specifically to active PSA. We used these to develop an immunopeptidometric assay (IPMA) that specifically detects this form of PSA. METHODS:Microtitration plates coated with a PSA antibody were used to capture PSA, and a PSA-binding glutathione S-transferase (GST) fusion peptide was used as a tracer. Bound tracer was detected with an antibody to GST labeled with a europium chelate. PSA isoenzymes with high and low enzymatic activity were used to study binding specificity. RESULTS:The IPMA detected enzymatically active PSA but not internally cleaved PSA and pro-PSA, which are enzymatically inactive. The assay detected 1-10% of free PSA in serum from PCa patients. CONCLUSIONS:Peptides identified by phage display can be used to develop assays with unique specificities for enzymatically active PSA. IPMA represents a new assay principle with wide potential utility.

journal_name

Clin Chem

journal_title

Clinical chemistry

authors

Wu P,Zhu L,Stenman UH,Leinonen J

doi

10.1373/clinchem.2003.026146

subject

Has Abstract

pub_date

2004-01-01 00:00:00

pages

125-9

issue

1

eissn

0009-9147

issn

1530-8561

pii

clinchem.2003.026146

journal_volume

50

pub_type

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