Surface Plasmon Resonance Imaging Analysis of Hexahistidine-tagged Protein on the Gold Thin Film Coated with a Calix Crown Derivative

• Published in: Biotechnology and bioprocess engineering Vol. 9; no. 2; pp. 143 - 146
• Main Authors: Chung, Bong-Hyun, Baek, Seung-Hak, Shin, Yong-Beom, Kim, Min-Gon, Ro, Hyeon-Su, Kim, Eun-Ki
• Format: Journal Article
• Language: Korean
• Published: 한국생물공학회 01.04.2004
• Subjects: 생물공학; surface plasmon resonance imaging (SPRI); calixcrown derivative; gold chip; hexahistidine-tagged protein
• ISSN: 1226-8372; 1976-3816

A surface plasmon resonance (SPR) imaging system was constructed and used to detect the hexahistidine-ubiquitin-tagged human parathyroid hormone fragment (His$\sub$6/-Ub-hPTHF(1-34)) expressed in Escherichia coli. The hexahistidine-specific antibody was immobilized on a thin gold film coated with ProLinker$\^$TM/ B, a novel calixcrown derivative with a bifunctional coupling property that permits efficient immobilizaton of capture proteins on solid matrices. The soluble and insoluble fractions of an E. coli cell lysate were spotted onto the antibody-coated gold chip, which was then washed with buffer (pH 7.4) solution and dried. SPR imaging measurements were carried out to detect the expressed His$\sub$6/-Ub-hPTHF(1-34). There was no discernible protein image in the uninduced cell lysate, indicating that non-specific binding of contaminant proteins did not occur on the gold chip surface. It is expected that the approach used here to detect affinity-tagged recombinant proteins using an SPR imaging technique could be used as a powerful tool for the analyses of a number of proteins in a high-throughput mode.