SPR imaging biosensor for aspartyl cathepsins: sensor development and application for biological material.

Abstract:

:A Surface Plasmon Resonance Imaging (SPRI) sensor has been developed for highly selective determination of cathepsin D (Cat D) or/and E (Cat E). The sensor contains immobilised pepstatin A, which binds aspartyl proteases from solution. Pepstatin A activated with N-Hydroxysuccinimide (NHS) and N-Ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) was immobilized on an amine-modified gold surface. Cysteamine was used for modification of the gold surface. Pepstatin A concentration and pH of interaction were optimised. A concentration of pepstatin equal to 0.5 microg mL(-1) and a pH of 3.75 were selected as optimal. The sensor's dynamic response range is between 0.25 and 1.0 ng mL(-1), and the detection limit is 0.12 ng mL(-1). However, the sensor cannot distinguish between Cat D and Cat E. In order to demonstrate the sensor's potential, Cat E was determined in human red blood cells, Cat D in human saliva, as well as total concentration of Cat D and Cat E in human nasal polyps.

journal_name

Protein Pept Lett

authors

Gorodkiewicz E,Regulska E

doi

10.2174/092986610791760450

subject

Has Abstract

pub_date

2010-09-01 00:00:00

pages

1148-54

issue

9

eissn

0929-8665

issn

1875-5305

pii

BSP/ PPL/ E pub/0137

journal_volume

17

pub_type

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