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Title 

Polyhydroxyalkanoate chip for the specific immobilization of recombinant proteins and its applications in immunodiagnostics

Authors 

T J ParkJ P ParkS J LeeHyo Jeong HongS Y Lee

Publisher 

Springer Verlag (Germany)

Issue Date 

2006

Citation 

Biotechnology and Bioprocess Engineering, vol. 11, no. 2, pp. 173-177

Keywords 

HBV preS2 surface proteinmicroarrayP(3HB) depolymerase substrate binding domainpoly(3-hydroxybutyrate)SARS-CoV envelope proteinhybrid proteinmembrane proteinrecombinant proteinvirus envelope proteinprotein domain

Abstract 

In this study, a novel strategy was developed for the highly selective immobilization of proteins, using the polyhydroxyalkanoate (PHA) depolymerase substrate binding domain (SBD) as an active binding domain. In order to determine the appropriacy of this method for immunodiagnostic assays, the single-chain antibody (ScFv) against the hepatitis B virus (HBV) preS2 surface protein and the severe acute respiratory syndrome coronavirus (SARS-CoV) envelope protein (SCVe) were fused to the SBD, then directly immobilized on PHA-coated slides via microspotting. The fluorescence-labeled HBV antigen and the antibody against SCVe were then utilized to examine specific interactions on the PHA-coated surfaces. Fluorescence signals were detected only at the spotted positions, thereby indicating a high degree of affinity and selectivity for their corresponding antigens/antibodies. Furthermore, we detected small amounts of ScFv-SBD (2.7 ng/mL) and SCVe-SBD fusion proteins (0.6 ng/mL). Therefore, this microarray platform technology, using PHA and SBD, appears generally appropriate for immunodiagnosis, with no special requirements with regard to synthetic or chemical modification of the biomolecules or the solid surface.

ISSN 

1226-8372

Link 

http://dx.doi.org/10.1007/BF02931904

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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