Methods for producing Anti-tnf antibody compositions

Abstract

Presented herein are a method for producing a recombinant anti-TNF antibody and a composition comprising the recombinant anti-TNF antibody, the recombinant anti-TNF antibody having a heavy chain (HC) comprising SEQ ID NO: 38 and a light chain (LC) comprising SEQ ID NO: 37.

Description

[Technical Field] Reference to the electronic submission sequence list This application contains a sequence list, which has been electronically submitted via EFS-Web in ASCII format. The file name is "JBI6053USPSP1SEQLIST.TXT", the creation date is December 7, 2018, and the file size is 25,153 bytes. The sequence list submitted via EFS-Web is an integral part of this specification, the entire text of which is incorporated herein by reference. This invention relates to a method for producing a recombinant anti-TNF antibody and a composition comprising the recombinant anti-TNF antibody having a heavy chain (HC) comprising the amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising the amino acid sequence SEQ ID NO: 37. [Previous Technology] TNFα is a soluble homotrimer of a 17kD protein subunit. A membrane-bound 26kD precursor form of TNF also exists. Cells other than monocytes or macrophages also produce TNFα. For example, human non-monocyte tumor cell lines produce TNFα and CD4+ and CD8+ peripheral blood T lymphocytes, and some cultured T and B cell lines also produce TNFα. TNFα induces pro-inflammatory action, which leads to tissue damage, such as degeneration of cartilage and bone, induces adhesion molecules, induces procoagulant activity on vascular endothelial cells, increases the adhesion of neutrophils and lymphocytes, and stimulates the release of platelet-activating factor from macrophages, neutrophils, and vascular endothelial cells. TNFα has been associated with infections, immune disorders, tumor pathology, autoimmune pathology, and graft-versus-host disease. The association of TNFα with cancer and infection pathology is often related to the host's catabolic state. Cancer patients often experience weight loss, which is frequently associated with anorexia. Extensive wasting associated with cancer and other diseases is called cachexia. Cachexia includes progressive weight loss, anorexia, and a persistent erosion of lean body mass in response to malignant growth. Cachexia contributes to higher cancer incidence and mortality. There is evidence that TNFα is involved in cachexia in cancer, infectious pathology, and other anaphylactic states. It is believed that TNFα plays a central role in Gram-negative sepsis and endotoxin-induced shock, including symptoms such as fever, malaise, anorexia, and cachexia. Endotoxin strongly activates monocyte / macrophage production and the secretion of TNFα and other cytokines. TNFα and other monocyte-derived cytokines mediate the metabolism of endotoxin and neurohormonal responses. Administration of endotoxin to human volunteers leads to acute illness with flu-like symptoms, including fever, tachycardia, increased metabolic rate, and release of stress hormones. Circulating TNFα is increased in patients with Gram-negative sepsis. Therefore, TNFα has been associated with inflammatory diseases, autoimmune diseases, viral, bacterial, and parasitic infections, malignant diseases, and / or neurodegenerative diseases, and is a useful target for specific biotherapies in diseases such as rheumatoid arthritis and Crohn's disease. Beneficial effects of TNFα monoclonal antibodies have been reported in open-label trials, including suppression of inflammation and successful retreatment after relapse of rheumatoid arthritis and Crohn's disease. Beneficial results have also been reported in randomized, double-blind, placebo-controlled trials of TNFα inhibition of inflammation in rheumatoid arthritis. In mammals other than humans, TNF-neutralizing antiserum or mAbs have been shown to eliminate adverse physiological changes and prevent death following lethal attacks of experimental endotoxemia and bacteremia. This effect has been confirmed, for example, in rodent lethality tests and primate pathological models. The putative receptor-binding locus of hTNF has been revealed, as has the receptor-binding locus of TNFα composed of amino acids 11 to 13, 37 to 42, 49 to 57, and 155 to 157 of INF. Non-human mammalian, chimeric, polyclonal (e.g., antiserum), and / or monoclonal antibodies (Mabs) and fragments (e.g., proteolytic digests or their fusion protein products) are potential therapeutic agents and have been investigated in some cases to attempt to treat certain diseases. However, such antibodies or fragments can induce an immune response when administered to humans. This immune response can lead to the clearance of the antibody or fragment from circulation mediated by immune complexes, making repeated administration unsuitable for therapy, thereby reducing the therapeutic benefit to the patient and limiting the administration of the antibody or fragment. For example, repeated administration of antibodies or fragments containing non-human portions can lead to serum sickness and / or severe anaphylaxis. To avoid these and other problems, many methods have been employed to reduce the immunogenicity of such antibodies and their portions, including chimerization and humanization, as well as are well known in the art. However, these and other methods may still result in antibodies or fragments exhibiting some immunogenicity, low affinity, low avidity, or problems in cell culture, scale-up, production, and / or low yield. Therefore, such antibodies or fragments may be less suitable for manufacturing or using as therapeutic proteins. Therefore, there is a need to provide anti-TNF antibodies or fragments thereof as therapeutic agents for treating diseases mediated by TNFα. [Summary of the Invention] Embodiments of the present invention are defined by the independent and supplementary claims appended herein, which are incorporated herein by reference for the sake of brevity. Other embodiments, features, and advantages of the various aspects of the invention will become apparent from the following description in conjunction with the accompanying drawings. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells). In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of such anti-TNF antibodies comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of such anti-TNF antibodies further comprises individual neutral oligosaccharide species G0F > 60.0%, G1F < 20.0%, and G2F < 5.0%. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies do not possess disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA). In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies are a follow-on biologic formulation. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of such anti-TNF antibodies comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species, and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of such anti-TNF antibodies further comprises individual neutral oligosaccharide species G0F > 60.0%, G1F < 20.0%, and G2F < 5.0%, and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies do not possess disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and such anti-TNF antibody systems are biosimilar formulations. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and such anti-TNF antibody systems are biosimilar formulations. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide form of the anti-TNF antibodies comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species, and wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells, and the anti-TNF antibody system is a biosimilar formulation. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID No: 37, wherein the anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of the anti-TNF antibodies further comprises individual neutral oligosaccharide species G0F > 60.0%, G1F < 20.0%, and G2F < 5.0%, and wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and the anti-TNF antibody system is a biosimilar formulation. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies do not possess disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and such anti-TNF antibody system is a biosimilar formulation. In some embodiments, the present invention provides anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein such anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein such anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and such anti-TNF antibody systems are biosimilar formulations. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies. In some embodiments, the present invention provides a method for producing anti-TNF antibodies, the anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the oligosaccharide form of the anti-TNF antibodies comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the oligosaccharide form of the anti-TNF antibodies further comprises individual neutral oligosaccharide species G0F > 60.0%, G1F < 20.0%, and G2F < 5.0%. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the anti-TNF antibodies do not contain disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA). In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the anti-TNF antibody system is a biosimilar formulation. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the oligosaccharide form of the anti-TNF antibodies comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species, and wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the oligosaccharide form of the anti-TNF antibodies further comprises individual neutral oligosaccharide species GOF > 60.0%, G1F < 20.0%, and G2F < 5.0%, and wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) containing nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the anti-TNF antibodies do not contain disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA), and wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides a method for producing anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies, wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and the anti-TNF antibody system is a biosimilar formulation. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells). In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide form of the anti-TNF antibody comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species. In some embodiments, the present invention provides a composition comprising anti-TNF antibodies, the anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of the anti-TNF antibodies further comprises individual neutral oligosaccharide species G0F > 60.0%, G1F < 20.0%, and G2F < 5.0%. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein, as determined by high performance liquid chromatography (HPLC), the anti-TNF antibody does not possess disialylated glycan species. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the anti-TNF antibody is a follow-on biologic. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain containing the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain containing the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of the anti-TNF antibody comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species, and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of the anti-TNF antibody further comprises individual neutral oligosaccharide species G0F > 60.0%, G1F < 20.0%, and G2F < 5.0%, and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells. In some embodiments, the present invention provides a composition comprising anti-TNF antibodies, the anti-TNF antibodies being: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibodies are expressed in Chinese hamster ovary cells (CHO cells), and wherein, as determined by high performance liquid chromatography (HPLC), the anti-TNF antibodies do not possess disialylated glycan species, and wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide form of the anti-TNF antibody comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species, and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells, and the anti-TNF antibody is a biosimilar formulation. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID No: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein the oligosaccharide-like form of the anti-TNF antibody further comprises individual neutral oligosaccharide species G0F>60.0%, G1F<20.0%, and G2F<5.0%, and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells, and the anti-TNF antibody is a biosimilar formulation. In some embodiments, the present invention provides a composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells), and wherein, as determined by high performance liquid chromatography (HPLC), the anti-TNF antibody does not possess disialylated glycan species, and wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / O cells, and the anti-TNF antibody is a biosimilar formulation. In some embodiments, the present invention provides an isolated nucleic acid molecule encoding a recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37. In some embodiments, the present invention provides a vector or recombinant eukaryotic host cell comprising a nucleic acid molecule encoding a recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising the amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising the amino acid sequence SEQ ID NO: 37. In some embodiments, the present invention provides a recombinant eukaryotic host cell comprising a nucleic acid molecule encoding a recombinant anti-TNF antibody comprising a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37, wherein the eukaryotic host cell line is selected from the group consisting of: COS-1 cells, COS-7 cells, HEK293 cells, BHK21 cells, Chinese hamster ovary (CHO) cells, BSC-1 cells, Hep G2 cells, P3X63Ag8.653(653) cells, Sp2 / 0 cells, HeLa cells, myeloma cells, and lymphoma cells. In some embodiments, the present invention provides a recombinant eukaryotic host cell comprising a nucleic acid molecule encoding a recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37, wherein the eukaryotic host cell line is an Sp2 / 0 cell. In some embodiments, the present invention provides a method for producing a recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising amino acid sequence SEQ ID NO: 37, the method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing an anti-TNF antibody molecule, the recombinant eukaryotic host cell comprising a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody. In some embodiments, the present invention provides a method for producing a recombinant anti-TNF antibody comprising: a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37, the method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing an anti-TNF antibody molecule, the recombinant eukaryotic host cell containing a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the eukaryotic host cell line is selected from the group consisting of: COS-1 cells, COS-7 cells, HEK293 cells, BHK21 cells, Chinese hamster ovary (CHO) cells, BSC-1 cells, Hep... G2 cells, P3X63Ag8.653(653) cells, Sp2 / 0 cells, HeLa cells, myeloma cells, and lymphoma cells. In some embodiments, the present invention provides a method for producing a recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising amino acid sequence SEQ ID NO: 37, the method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing an anti-TNF antibody molecule, the recombinant eukaryotic host cell comprising a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the eukaryotic host cell line is Sp2 / 0 cells. In some embodiments, the present invention provides a recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody system is produced by a method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing an anti-TNF antibody molecule, the recombinant eukaryotic host cell comprising a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody. In some embodiments, the present invention provides a recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising the amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising the amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody system is produced by a method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing an anti-TNF antibody molecule, the recombinant eukaryotic host cell comprising a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising the amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising the amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the anti-TNF antibody inhibits the activity of TNFα. In some embodiments, the present invention provides a recombinant anti-TNF antibody comprising: a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody system is produced by a method comprising: a. culturing the recombinant eukaryotic host cells under conditions expressing anti-TNF antibody molecules, the recombinant eukaryotic host cells containing nucleic acid molecules encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the eukaryotic host cell line is selected from the group consisting of: COS-1 cells, COS-7 cells, HEK293 cells, BHK21 cells, Chinese hamster ovary (CHO) cells, BSC-1 cells, Hep... G2 cells, P3X63Ag8.653(653) cells, Sp2 / 0 cells, HeLa cells, myeloma cells, and lymphoma cells. In some embodiments, the present invention provides a recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising the amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising the amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody system is produced by a method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing an anti-TNF antibody molecule, the recombinant eukaryotic host cell comprising a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising: a heavy chain (HC) comprising the amino acid sequence SEQ ID NO: 38 and a light chain (LC) comprising the amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the eukaryotic host cell line is Sp2 / 0 cells. In some embodiments, the present invention provides a pharmaceutical composition comprising a recombinant anti-TNF antibody comprising a heavy chain (HC) containing amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody system is produced by a method comprising: a. culturing the recombinant eukaryotic host cell containing a nucleic acid molecule encoding the recombinant anti-TNF antibody under conditions expressing the anti-TNF antibody molecule, the recombinant anti-TNF antibody comprising a heavy chain (HC) containing amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody. In some embodiments, the present invention provides a pharmaceutical composition comprising a recombinant anti-TNF antibody comprising a heavy chain (HC) containing amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody is produced by a method comprising: a. culturing the recombinant eukaryotic host cell containing a nucleic acid molecule encoding the recombinant anti-TNF antibody under conditions expressing the anti-TNF antibody molecule, the recombinant anti-TNF antibody comprising a heavy chain (HC) containing amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the anti-TNF antibody inhibits TNFα activity. In some embodiments, the present invention provides a pharmaceutical composition comprising a recombinant anti-TNF antibody comprising a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody is produced by a method comprising: a. culturing the recombinant eukaryotic host cells under conditions expressing the anti-TNF antibody molecule, the recombinant eukaryotic host cells comprising a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising a heavy chain (HC) containing the amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing the amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the eukaryotic host cell line is selected from the group consisting of: COS-1 cells, COS-7 cells, HEK293 cells, BHK21 cells, Chinese hamster ovary (CHO) cells, BSC-1 cells, Hep... G2 cells, P3X63Ag8.653(653) cells, Sp2 / 0 cells, HeLa cells, myeloma cells, and lymphoma cells. In some embodiments, the present invention provides a pharmaceutical composition comprising a recombinant anti-TNF antibody comprising a heavy chain (HC) containing amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing amino acid sequence SEQ ID NO: 37, wherein the recombinant anti-TNF antibody system is produced by a method comprising: a. culturing the recombinant eukaryotic host cell under conditions expressing the anti-TNF antibody molecule, the recombinant eukaryotic host cell containing a nucleic acid molecule encoding the recombinant anti-TNF antibody, the recombinant anti-TNF antibody comprising a heavy chain (HC) containing amino acid sequence SEQ ID NO: 38 and a light chain (LC) containing amino acid sequence SEQ ID NO: 37; and b. recovering the recombinant anti-TNF antibody, wherein the eukaryotic host cell line is Sp2 / 0 cells. [Simplified Explanation of the Diagram] [Figure 1] shows a graphical representation of the assay for the ability of TNV mAb to inhibit the binding of TNFα to the recombinant TNF receptor in fusion tumor cell supernatant. Different amounts of fusion tumor cell supernatant containing known amounts of TNV mAb were pre-cultured with a fixed concentration (5 ng / ml) of 125I-labeled TNFα. The mixture was transferred to 96-well Optiplates pre-coated with p55-sf2 (a recombinant TNF receptor / IgG fusion protein). After washing away unbound material, the amount of TNFα bound to the p55 receptor in the presence of the mAb was determined and counted using a γ-counter. Although eight TNV ​​mAb samples were tested in these experiments, for simplicity, the three mAbs that were identified by DNA sequence analysis as being identical to one of the other TNV mAbs are not shown here (see Section 5.2.2). Each sample was tested repeatedly. The results shown represent two independent experiments. Figures 2A and 2B show the DNA sequence of the variable region of the TNV mAb heavy chain. The germline gene shown is the DP-46 gene. The "TNV" indicator shows the sequences TNV14, TNV15, TNV148, and TNV196. The first three nucleotides in the TNV sequence define the translation start Met codon. The dots in the TNV mAb gene sequence indicate nucleotides that are the same as those in the germline sequence. The first 19 nucleotides (underlined) of the TNV sequence correspond to the oligonucleotides used for PCR amplification of the variable region. Only for the germline gene are amino acid translations (single-letter abbreviations) starting with mature mAb shown. The three CDR domains in the germline amino acid translation are marked in bold and underlined. The row indicating TNV148(B) indicates the sequence for both TNV148 and TNV148B. The gaps in the germline DNA sequence (CDR3) are because the sequence is unknown or absent in the germline gene. The TNV mAb heavy chain uses the J6 joining region. [Figure 3] shows the DNA sequence of the variable region of the TNV mAb light chain. The germline gene shown is a representative member of the Vg / 38K family of human κ germline variable region genes. The dotted nucleotides in the TNV mAb gene sequence are identical to those in the germline sequence. The first 16 nucleotides (underlined) of the TNV sequence correspond to the oligonucleotides used for PCR amplification of the variable region. Only the amino acid translation of the mature mAb (single-letter abbreviation) is shown for the germline gene. The three CDR domains in the germline amino acid translation are marked in bold and underlined. The row indicating TNV148(B) indicates the sequence for both TNV148 and TNV148B. The gaps in the germline DNA sequence (CDR3) are because the sequence is unknown or absent in the germline gene. The TNV mAb light chain uses the J3 joining sequence. [Figure 4] shows the deduced amino acid sequences of the variable region of the heavy chain of TNV mAb. The amino acid sequences shown (single-letter abbreviations) are deduced from the DNA sequences determined by both unselected and selected PCR products. The amino acid sequences shown are divided into secretion signal sequence (signal), fabrication layer (FW), and complementarity-determining region (CDR) domains. The amino acid sequences of the DP-46 germline gene are shown in the top row of each domain. Dots indicate that the amino acids in the TNV mAb are the same as those in the germline gene. TNV148(B) indicates that the sequence shown is for both TNV148 and TNV148B. "TNV" indicates that the sequence shown is for all TNV mAbs unless a different sequence is shown. The dash in the germline sequence (CDR3) indicates that such sequences are unknown or absent in the germline gene. [Figure 5] shows the deduced amino acid sequences of the variable regions of the TNV mAb light chain. The amino acid sequences shown (single-letter abbreviations) are deduced from the DNA sequences determined by both unselected and selected PCR products. The amino acid sequences shown are divided into secretion signal (signal), fabrication (FW), and complementarity-determining region (CDR) domains. The amino acid sequences of the Vg / 38K type light chain germline gene are shown in the top row of each domain. Dots indicate that the amino acids in TNV mAb are the same as those in the germline gene. TNV148(B) indicates that the sequence shown pertains to both TNV148 and TNV148B. "All" indicates that the sequences shown pertain to TNV14, TNV15, TNV148, TNV148B, and TNV186. [Figure 6] shows a schematic diagram of the heavy and light chain expression plastids used to create C466 cells expressing rTNV148B. p1783 is the heavy chain plastid and p1776 is the light chain plastid. The variable and constant region coding domains of rTNV148B are shown in black boxes. The immunoglobulin enhancers in the JC intron are shown in gray boxes. Relevant restriction sites are shown. These plastids are shown oriented so that transcription of the Ab gene proceeds in a clockwise direction. The length of plastid p1783 is 19.53 kb and the length of plastid p1776 is 15.06 kb. The complete nucleotide sequences of both plastids are known. By substituting the BsiWI / BstBI restriction fragment, the variable region coding sequence in p1783 can be easily replaced with another heavy chain variable region sequence. By substituting the SalI / AflII restriction fragment, the variable region coding sequence in p1776 can be replaced with another variable region sequence. [Figure 7] shows a graphical representation of the growth curve analysis for five rTNV148B cell lines. Cells were seeded into T75 flasks in I5Q+MHX medium to achieve a viable cell density of 1.0 × 10⁵ cells / ml in a 30 ml volume, starting on day 0. Due to transfection and subcloning, the cell cultures used in these studies were continuously cultured. Over the following days, the cells in the T flasks were completely resuspended, and 0.3 ml aliquots of culture were removed. Growth curve studies were terminated when the cell count dropped below 1.5 × 10⁵ cells / ml. The number of viable cells in the aliquots was determined using trypan blue exclusion, and the remaining portions of the aliquots were stored for later mAb concentration determination. Simultaneously, all sample aliquots were subjected to an ELISA for human IgG. [Figure 8] A graphical representation showing the comparison of cell growth rates with different concentrations of MHX. Secondary cell lines C466A and C466B were thawed into MHX-free medium (IMDM, 5% FBS, 2mM glutamic acid) and cultured for an additional two days. The two cell cultures were then divided into three cultures: one without MHX, one containing 0.2X MHX, and one containing 1X MHX. One day later, fresh T75 flasks were seeded with an initial density of 1×10⁵ cells / ml, and cell counts were performed at 24-hour intervals for one week. The doubling time during the first 5 days was calculated using the formula in SOP PD32.025 and is shown above the bars. [Figure 9] shows a graphical representation of the mAb production stability over time for two cell lines producing rTNV148B. Secondary cell lines that had been continuously cultured since transfection and secondary selection were used to initiate long-term continuous culture in 24-well dishes. Cells were cultured in I5Q medium with and without MHX selection. Cells were continuously subcultured by isolating cultures every 4 to 6 days to maintain new live cultures while rendering previous cultures ineffective. Aliquots of used cell supernatant were collected shortly after culture use and stored until mAb concentration was determined. All aliquots were simultaneously subjected to an ELISA against human IgG. [Figure 10] shows the weight change of Tg 197 mice in an arthritis mouse model responding to the anti-TNF antibody of the present invention compared to the control group in Example 4. At approximately 4 weeks of age, Tg197 mice were assigned to one of nine treatment groups based on sex and body weight and treated with a single intraperitoneal bolus dose of 1 mg / kg or 10 mg / kg of Dulbecco's PBS (D-PBS) or the anti-TNF antibody of the present invention (TNV14, TNV148, or TNV196). When analyzing weight changes from pre-treatment, the weight gain of animals treated with 10 mg / kg cA2 was consistently greater than that of animals treated with D-PBS throughout the study. This weight gain was significant from week 3 to week 7. Animals treated with 10 mg / kg TNV148 also achieved a significant weight gain at week 7 of the study. Figures 11A to 11C illustrate the progression of disease severity based on the arthritis index as presented in Example 4. The arthritis index of the group treated with 10 mg / kg cA2 was lower than that of the D-PBS control group, starting from week 3 and continuing into the remainder of the study (week 7). Compared to the D-PBS group, neither the animals treated with 1 mg / kg TNV14 nor those treated with 1 mg / kg cA2 showed a significant reduction in AI after week 3. There were no significant differences between the 10 mg / kg treatment groups when compared to other similar doses (10 mg / kg cA2 vs. 10 mg / kg TNV14, 148, and 196). When comparing the 1 mg / kg treatment groups, 1 mg / kg TNV148 showed significantly lower AI than 1 mg / kg cA2 at weeks 3, 4, and 7. At weeks 3 and 4, 1 mg / kg TNV148 was also significantly lower than the group treated with 1 mg / kg TNV14. Although TNV196 showed a significant reduction in AI up to week 6 of the study (compared to the D-PBS-treated group), TNV148 was the only treatment that maintained a significant 1 mg / kg reduction at the end of the study. [Figure 12] shows the weight change of Tg 197 mice in an arthritis mouse model responding to the anti-TNF antibody of the present invention compared to the control group in Example 5. At approximately 4 weeks of age, Tg197 mice were assigned to one of eight treatment groups based on body weight and treated with either the control group material (D-PBS) or the antibody (TNV14, TNV148) via intraperitoneal bolus injection at 3 mg / kg (week 0). Repeat injections were administered to all animals at weeks 1, 2, 3, and 4. The efficacy of the test material in groups 1 through 6 was assessed. Immune response induction and pharmacokinetic clearance of TNV14 or TNV148 in serum samples obtained from animals in groups 7 and 8 were assessed at weeks 2, 3, and 4. Figures 13A to 13C illustrate the progression of disease severity in Example 5 based on the arthritis index. The arthritis index in the group treated with 10 mg / kg cA2 was significantly lower than that in the D-PBS control group, starting from week 2 and continuing into the remainder of the study (week 5). Compared to the D-PBS control group, animals treated with 1 mg / kg or 3 mg / kg cA2, and animals treated with 3 mg / kg TNV14, did not achieve any significant reduction in AI at any point during the study. Compared to the D-PBS group, animals treated with 3 mg / kg TNV14 showed a significant reduction, starting from week 3 and continuing into week 5. Compared to the lower doses of cA2 (1 mg / kg and 3 mg / kg) at weeks 4 and 5 of the study, animals treated with 10 mg / kg cA2 showed a significant reduction in AI, and were also significantly lower than those treated with TNV14 from weeks 3 to 5. Although there appeared to be no significant difference between any of the 3 mg / kg treatment groups, the AI ​​of animals treated with 3 mg / kg TNV14 was significantly higher than that of animals treated with 10 mg / kg at some time points, while animals treated with TNV148 were not significantly different from those treated with 10 mg / kg cA2. [Figure 14] shows the weight change of Tg 197 mice in an arthritis mouse model responding to the anti-TNF antibody of the present invention compared to the control group in Example 6. At approximately 4 weeks of age, Tg197 mice were assigned to one of six treatment groups based on sex and body weight, and treated with a single intraperitoneal bolus dose of antibody (cA2 or TNV148) of 3 mg / kg or 5 mg / kg. D-PBS and a 10 mg / kg cA2 control group were used in this study. [Figure 15] illustrates the progression of disease severity based on the Arthritis Index as presented in Example 6. All treatment groups showed some protective effect at earlier time points, with 5 mg / kg cA2 and 5 mg / kg TNV148 showing significant reductions in AI at weeks 1–3, and all treatment groups showing significant reductions at week 2. Later in the study, animals treated with 5 mg / kg cA2 showed some protective effect, with significant reductions at weeks 4, 6, and 7. Low doses of both cA2 and TNV148 (3 mg / kg) showed significant reductions at week 6, and all treatment groups showed significant reductions at week 7. No treatment group was able to maintain a significant reduction at the end of the study (week 8). There were no significant differences between any treatment groups (excluding the saline control group) at any time point. [Figure 16] shows the weight change of Tg 197 mice in an arthritis mouse model responding to the anti-TNF antibody of the present invention compared to the control group in Example 7. To compare the efficacy of a single intraperitoneal dose of TNV148 (derived from fusion tumor cells) versus rTNV148B (derived from transfected cells), Tg197 mice were assigned to one of nine treatment groups based on sex and body weight at approximately 4 weeks of age and treated with a single intraperitoneal bolus dose of 1 mg / kg of Dalberks PBS (D-PBS) or the antibody (TNV148, rTNV148B). [Figure 17] illustrates the progression of disease severity based on the arthritis index as presented in Example 7. The arthritis index of the group treated with 10 mg / kg cA2 was lower than that of the D-PBS control group, starting from week 4 and continuing into the remainder of the study (week 8). Both the TNV148-treated group and the 1 mg / kg cA2-treated group showed a significant reduction in AI at week 4. Although a previous study (P-099-017) showed that TNV148 was slightly more effective in reducing the arthritis index after a single intraperitoneal bolus of 1 mg / kg, this study showed that the AI ​​was slightly higher in both versions of the TNV antibody-treated group. Although (except at week 6) the 1 mg / kg cA2 group did not show a significant improvement compared to the 10 mg / kg cA2 group, and the TNV148 group had a significantly higher improvement at weeks 7 and 8, there was no significant difference in AI at any point in the study among 1 mg / kg cA2, 1 mg / kg TNV148, and 1 mg / kg TNV148B. [Figure 18] shows an overview of the nine stages of the golimumab manufacturing process. [Figure 19] shows a flowchart of the first-stage manufacturing process for the pre-culture and amplification steps, which includes process control and process monitoring tests. [Figure 20] shows the flowchart of the second stage manufacturing process, which includes process control and process monitoring tests. [Figure 21] A schematic overview of some of the major N-linked oligosaccharide species in golimumab IgG. It also shows the roles of some enzymes in the glycosylation maturation process and the roles of some divalent cations (e.g., Mn2+ as a cofactor and Cu2+ as an inhibitor of GalTI) (see, for example, Biotechnol Bioeng. 2007 Feb 15; 96(3): 538-49; Curr Drug Targets. 2008 Apr; 9(4): 292-309; J Biochem Mol Biol. 2002 May 31; 35(3): 330-6). It should be noted that the species with terminal sialic acid (S1 and S2) are charged species, while the species lacking terminal sialic acid (G0F, G1F, and G2F) are neutral species. However, the generation of charged species depends on the presence of galactose in G1F and G2F, which are added by the GalT1 enzyme. [Figure 22] shows representative HPLC chromatograms of oligosaccharide analysis of golimumab reference standards using HPLC and fluorescence detection. Peaks associated with different species are indicated. * indicates systematic peaks not associated with golimumab. [Figure 23] Representative deconvoluted mass spectra of IRMA analysis of golimumab produced in Sp2 / 0 cells. [Figure 24] shows the aspartic acid reaction mediated by cycloimide in the protein; the figure is modified from (Voorter, CE, et al. (1988). "Spontaneous peptide bond cleavage in aging alpha-crystallin through a succinimide intermediate." J Biol Chem 263 (35): 19020-19023). [Figure 25] shows the representative Lys C peptide chromatogram (214 nm) of golimumab. Modified (gray) and parental (black) peptides of interest are indicated. The peak at approximately 34 min is not associated with golimumab. [Figure 26] shows the peptiform spectra of golimumab peptides after 0, 4, 8, and 24 hours under forced deamination conditions, illustrating the changes in the levels of peptides 1-58 and 1-59 (left panel). This change is due to the deamination of Asn43 to Asp43 and isoAsp43, and the isomerization of isoAsp43 to Asp43 (top right panel; abundance was calculated using the relative peak areas of the Asn, Asp, and isoAsp forms of peptides 1-58 and 1-59). The Asn43, Asp43, and isoAsp43 forms of peptides 1-58 and 1-59 were identified by mass spectrometry (MS) (bottom right panel) and by comparison with the residence time of the synthesized Asp43 peptide 1-58 (not shown). [Figure 27] shows a representative cIEF electrophoresis profile of golimumab with four major peaks labeled C, 1, 2, and 3, and one minor peak labeled B. Internal standards of pI 7.6 and 9.5 are also indicated. [Figure 28] shows the correlation analysis between the efficacy of golimumab and LC cycAsn93%. The LC cycAsn93 percentage was determined using a deamination assay. Results are shown at 49 time points from different batches and quantities of LC cycAsn93. Statistical analysis was performed using commercial software; r is Pearson's correlation coefficient.

Implementation Method

Claims

1. An anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells).

2. The anti-TNF antibody as described in claim 1, wherein the oligosaccharide form of the anti-TNF antibody comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species.

3. The anti-TNF antibody as claimed in claim 1, wherein the oligosaccharide form of the anti-TNF antibody further comprises individual neutral oligosaccharide species G0F>60.0%, G1F<20.0%, and G2F<5.0%.

4. The anti-TNF antibody as claimed in claim 1, wherein the anti-TNF antibody does not contain disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA).

5. An anti-TNF antibody as described in any one of claims 1 to 4, wherein the anti-TNF antibody has a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells.

6. The anti-TNF antibody as described in claim 5, wherein the anti-TNF antibody system is a biosimilar formulation (follow-on biologic).

7. The recombinant anti-TNF antibody as described in claim 5, wherein the anti-TNF antibody inhibits the activity of TNFα.

8. A pharmaceutical composition comprising the recombinant anti-TNF antibody as described in claim 5.

9. A method for producing anti-TNF antibodies, the anti-TNF antibodies comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody system is produced by a method comprising: a. culturing Chinese hamster ovary cells (CHO cells) having nucleotides encoding the anti-TNF antibodies; b. expressing the anti-TNF antibodies in the CHO cells; and c. purifying the anti-TNF antibodies.

10. The manufacturing method as described in claim 7, wherein the oligosaccharide samples of the anti-TNF antibodies comprise >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species, and 11. The manufacturing method as described in claim 7, wherein the oligosaccharide samples of the anti-TNF antibodies further comprise individual neutral oligosaccharide species G0F>60.0%, G1F<20.0%, and G2F<5.0%.

12. The manufacturing method as described in claim 7, wherein the anti-TNF antibodies do not contain disialylated glycan species as determined by high performance liquid chromatography (HPLC) or reduced mass analysis (RMA).

13. The manufacturing method as described in any one of claims 7 to 10, wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells.

14. The manufacturing method as described in claim 11, wherein the anti-TNF anti-system biosimilar formulations are...

15. A composition comprising an anti-TNF antibody, the anti-TNF antibody comprising: (i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 38; and (ii) a light chain comprising the amino acid sequence of SEQ ID NO: 37, wherein the anti-TNF antibody is expressed in Chinese hamster ovary cells (CHO cells).

16. The composition as described in claim 13, wherein the oligosaccharide form of the anti-TNF antibodies comprises >99.0% total neutral oligosaccharide species and <1.0% total charged oligosaccharide species.

17. The composition as described in claim 13, wherein the oligosaccharide form of the anti-TNF antibodies further comprises individual neutral oligosaccharide species G0F>60.0%, G1F<20.0%, and G2F<5.0%.

18. The composition as claimed in claim 13, wherein, as determined by high performance liquid chromatography (HPLC), the anti-TNF antibodies do not contain disialylated polysaccharide species.

19. A composition as claimed in any one of claims 13 to 16, wherein the anti-TNF antibodies have a longer half-life or increased antibody-dependent cell-mediated cytotoxicity (ADCC) compared to anti-TNF antibodies expressed in Sp2 / 0 cells.

20. The composition as described in claim 17, wherein the anti-TNF anti-system biosimilar formulation.