發明
中華民國
106123547
I 777963
縱排重複性抗體結合蛋白及其應用
臺北醫學大學
2022/09/21
本發明利用基因工程技術,將可專一性結合抗體Fc (fragment-crystllizable)區域之protein G建構成串聯式重複蛋白質(poly-protein G)。Poly-protein G抓取抗體之效率與數量遠優於傳統單一protein G。Poly-protein G可固定於固相平台(solid phase plate),提升固相平台表面之抗體乘載量,並使抗體呈現之方向性一致;poly-protein G之另一應用,可直接與抗體混合,形成poly-protein G/抗體複合物,大幅提升抗體聚集於抗原區域之數量。本發明之另一應用係將poly-protein G表現於細胞表面,形成poly-protein G細胞;將poly-protein G細胞固定於固相平台,提升固相平台表面之抗體乘載量,並使抗體呈現之方向性一致;poly-protein G細胞之另一應用,可直接與抗體混合,形成poly-protein G細胞/抗體複合物,大幅提升抗體聚集於抗原區域之數量。與poly-protein G或poly-protein G細胞搭配之抗體可不須純化,即可使用於免疫測定法。本發明能大幅提升各種抗體相關免疫測定法之偵測敏感度與極限值。 The present invention relates to a recombinant protein, named poly-protein G, which is constructed from the fragment crystallisable (Fc)-binding domain of protein G by gene engineering technologies. Poly-protein G exhibits a much greater binding efficiency and affinity to antibody than monomer protein G does. By coating poly-protein G on a solid phase plate (e.g. plate, membrane and bead), the antibody-loading amount on the plate can significantly increase and the display of antigen binding domain (Fab) of antibody on the plate can be unidirectional (outward). Another application of poly-protein G is to generate poly-protein G/antibody complexes by simply mixing poly-protein G with antibodies, and increases the accumulated amount of the antibodies on the antigen area. Moreover, the invention relates to a poly-protein G cell, which expresses poly-protein G on cell surface. By coating poly-protein G cells on a solid phase plate, the antibody-loading amount on the plate can significantly increase and the display of antigen binding domain (Fab) of antibody on the plate can be unidirectional (outward). Another application of poly-protein G cells is to generate poly-protein G cell/antibody complexes by simply mixing the cells with antibodies, and increases the accumulated amount of the antibodies on the antigen area. Pairing with poly-protein G or poly-protein G cells allows antibodies directly appling to immunoassays without additional puritification. The invention can dramatically improve the sensitivities and detection limits of immunoassays using various antibodies.
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