His-Fc-G67 Polypeptide Production

His-Fc-G67 polypeptide was prepared by our previous methods with some modifications [36 (link)]. Fc-G67 contains the amino acid sequence derived from C1 region of Protein G, which strongly binds to IgG Fc region. A DNA fragment encoding the forward linker, C1 region, and the backward linker cysteine was synthesized by FASMAC Co., Ltd. (Kanagawa, Japan) and was then cloned into pET302/NT-His (Thermo Fisher Scientific, Waltham, MA, USA). The recombinant proteins were produced in BL21(DE3) cells (Thermo Fisher Scientific) using MagicMedia^TM E coli expression medium (Thermo Fisher Scientific) and were purified with TALON Metal Affinity Resin (Takara Bio Inc., Shiga, Japan).

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Yano Y., Hamano N., Haruta K., Kobayashi T., Sato M., Kikkawa Y., Endo-Takahashi Y., Tada R., Suzuki R., Maruyama K., Nomizu M, & Negishi Y. (2022). Development of an Antibody Delivery Method for Cancer Treatment by Combining Ultrasound with Therapeutic Antibody-Modified Nanobubbles Using Fc-Binding Polypeptide. Pharmaceutics, 15(1), 130.

Publication 2022

BL21(DE3) cells TALON Metal Affinity Resin

A dna Amino acid sequence Cells Cysteine E coli Metal Polypeptide Protein region Recombinant proteins Resin Talon

Corresponding Organization : Tokyo University of Pharmacy and Life Sciences

Other organizations : Teikyo University

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Variable analysis

independent variables

Modifications to the previous methods for preparing the His-Fc-G67 polypeptide

dependent variables

Production and purification of the recombinant His-Fc-G67 polypeptide

control variables

Amino acid sequence derived from the C1 region of Protein G, which strongly binds to the IgG Fc region
Forward linker, C1 region, and backward linker cysteine DNA fragment cloned into pET302/NT-His vector
Recombinant protein production in BL21(DE3) cells using MagicMedia E coli expression medium
Purification of recombinant proteins using TALON Metal Affinity Resin

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