METHOD FOR PLACING ANTIBODIES IN THE CYTOPLASMA IN THE FORM OF FULL IMMUNOGLOBULIN BY PERMISSION OF ANTIBODIES THROUGH A CELLULAR MEMBRANE AND ITS APPLICATION

摘要

The present invention relates to a method of localizing an intact immunoglobulin-format antibody in cytosol by permeating membrane of cells. The present invention also relates to a light-chain variable region (VL) that induces an intact immunoglobulin-format antibody to penetrate the membrane of living cells and be localized in the cytosol, and to an antibody comprising the same. The present invention also relates to a biologically active molecule fused to the antibody and selected from the group consisting of peptides, proteins, small-molecule drugs, nanoparticles and liposomes. The present invention also relates to a composition for prevention, treatment or diagnosis of cancer, comprising: the antibody; or a biological active molecule fused to the antibody and selected from the group consisting of peptides, proteins, small-molecule drugs, nanoparticles and liposomes. The present invention also relates to a polynucleotide that encodes the light-chain variable region and the antibody. The present invention also relates to a method for producing an antibody which penetrates cells and is localized in the cytosol. According to the method of the present invention, which allows an intact immunoglobulin-format antibody to actively penetrate living cells and be localized in the cytosol, the antibody can penetrate living cells and be localized in the cytosol, without having to use a special external protein delivery system. Moreover, the use of the cytosol-penetrating light-chain variable region according to the present invention and the intact immunoglobulin-format antibody comprising the same can penetrate cells and remain in the cytosol, without affecting the high specificity and affinity of a human antibody heavy-chain variable region (VH) for antigens, and thus can be localized in the cytosol which is currently classified as a target in disease treatment based on small-molecule drugs, and at the same time, can exhibit high effects on the treatment and diagnosis of tumor and disease-related factors that show structurally complex interactions through a wide and flat surface between protein and protein. In addition, these can selectively inhibit KRas mutants, which are major drug resistance-associated factors in the use of various conventional tumor therapeutic agents, and at the same time, can be used in combination with conventional therapeutic agents to thereby exhibit effective anticancer activity.