Difference between revisions of "Part:BBa K2885002"

Revision as of 18:16, 16 October 2018

Gold binding polypeptide (GBP) + Protein G (ProG) Fusion

Gold binding polypeptide (GBP)-Protein G (ProG)(GBP-ProG) in our study was constructed from Integrated DNA Technologies (IDT) and the sequence was analyzed by BIOFACTTM sequencing analysis service. As we mentioned properties of GBP and ProG in separate parts of GBP (BBa_K2885001) and ProG (BBa_K2885000), this part covered our fusion protein’s function as a crosslinker for antibody immobilization on a gold surface.

To shed light on successful immobilization of antibody by the GBP-ProG on the gold surface, we applied the GBP-ProG to the gold surface in the microchannel of our chip, followed by injection of the alkaline phosphatase (AP)-labeled immunoglobulin G (IgG) to immobilize onto the GBP-ProG formed. AP-labeled IgG also was loaded on a bare gold surface inside the microchannel as a control. Then PAPP was treated into both microchannels to observe cyclic voltammograms (CV) induced by its enzymatic reaction as a substrate. CV current values were 38.63 μA at conditions in which GBP-ProG exists in spite of no cyclic voltammetric peak responses in without GBP-ProG. This result showed that GBP-ProG was successfully immobilized onto the gold surface via the GBP portion, indicating the oriented binding of antibodies onto the ProG domain targeting the Fc region of antibodies. Hence, we clarified that our GBP-ProG can be an efficient crosslinker of diverse antibodies.

Sequence and Features