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本開示は一般用語で記載されてきたが、本開示の実施形態は、特許請求の範囲の範囲を限定するものと解釈されるべきではない以下の実施例で更に開示される。
本発明の様々な実施形態を以下に示す。
１．約１５％～約２７％のＧ１Ｆオリゴ糖含有量を有する、配列番号７の重鎖可変領域（ＶＨ）及び配列番号８の軽鎖可変領域（ＶＬ）をコードしているポリヌクレオチドから発現する抗ＣＤ３８抗体の生成方法であって、
ａ）約８．５十億分率（ｐｐｂ）以下のマンガン（Ｍｎ）を含む培養培地を調製することと、
ｂ）工程ａ）で調製された前記培養培地中で前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドを含む宿主細胞を培養することによって、前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量を制御し、それにより、前記約１５％～約２７％のＧ１Ｆオリゴ糖含有量を有する、前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドから発現する前記抗ＣＤ３８抗体を生成することと、
を含む方法。
２．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が、約１５％～約２５％である、上記１に記載の方法。
３．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が、約２１％～約２５％である、上記１に記載の方法。
４．前記抗ＣＤ３８抗体のＧ０Ｆオリゴ糖含有量が、約６５％～約７４％である、上記１～３のいずれかに記載の方法。
５．前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が、約６８％～７４％である、上記１～３のいずれかに記載の方法。
６．前記培養培地を調製することが、前記培養培地を調製するために使用される原材料成分の１つ以上のバッチにおけるＭｎ濃度を測定することと、組み合わせて約８．５ｐｐｂ以下のＭｎを含有する原材料成分の１つ以上のバッチを選択することと、を含む、上記１～５のいずれかに記載の方法。
７．前記培養培地が、約４．０ｐｐｂ～約８．５ｐｐｂのＭｎを含むように調製される、上記６に記載の方法。
８．前記培養培地が、約４．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記７に記載の方法。
９．前記培養培地が、約５．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記８に記載の方法。
１０．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約１５％～約２７％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６５％～約７４％であり、前記培養培地が、約４．０ｐｐｂ～約８．５ｐｐｂのＭｎを含むように調製される、上記１～９のいずれかに記載の方法。
１１．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約１５％～約２５％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６８％～約７４％であり、前記培養培地が、約４．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記１～９のいずれかに記載の方法。
１２．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約２１％～約２５％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６８％～約７４％であり、前記培養培地が、約５．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記１～９のいずれかに記載の方法。
１３．前記培養培地が、基本培地又は流加培地である、上記１～１２のいずれかに記載の方法。
１４．培養が、流加培養又は灌流培養を含む、上記１～１３のいずれかに記載の方法。
１５．前記宿主細胞が、真核細胞である、上記１～１４のいずれかに記載の方法。
１６．前記真核細胞が、ＣＨＯ細胞、ＰＥＲ．Ｃ６細胞、ＮＳ０細胞、Ｓｐ２／０細胞、又はＢＨＫ細胞である、上記１６に記載の方法。
１７．前記ＣＨＯ細胞が、ＣＨＯ－Ｋ１細胞、ＣＨＯ－ＤＧ４４細胞、ＣＨＯ－Ｓ細胞、又はＣＨＯ－ＤＸＢ１１細胞である、上記１６に記載の方法。
１８．前記ＣＨＯ細胞が、グルタミン合成酵素（ＧＳ）が不足している、上記１７に記載の方法。
１９．ＧＭＰに準拠する条件下で実施される、上記１～１８のいずれかに記載の方法。
２０．前記抗ＣＤ３８抗体が、前記配列番号７のＶＨ及び前記配列番号９のＶＬを含む、上記１～１９のいずれかに記載の方法。
２１．前記抗ＣＤ３８抗体が、ＩｇＧ１アイソタイプを含む、上記１～２０のいずれかに記載の方法。
２２．前記抗ＣＤ３８抗体が、配列番号９の重鎖（ＨＣ）及び配列番号１０の軽鎖（ＬＣ）を含む、上記１～２１のいずれかに記載の方法。
２３．前記抗ＣＤ３８抗体が、バイオ後続品である、上記１～２２のいずれかに記載の方法。
２４．約１５％～約２７％のＧ１Ｆオリゴ糖含有量を有する、配列番号７のＶＨ及び配列番号８のＶＬをコードしているポリヌクレオチドから発現する抗ＣＤ３８抗体の生成方法であって、
ａ）前記抗ＣＤ３８抗体が生成される条件下で前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドを発現する宿主細胞を培養することと、
ｂ）前記抗ＣＤ３８抗体の生合成中の培養培地におけるＭｎの濃度をモニタリングし、前記抗ＣＤ３８抗体の生合成中の前記培養培地におけるＭｎの濃度を調節することによって、前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量を制御し、前記培養培地におけるＭｎの濃度を約８．５ｐｐｂ以下のＭｎを含むように調節し、それにより、前記約１５％～約２７％のＧ１Ｆオリゴ糖含有量を有する前記抗ＣＤ３８抗体を生成することと、
を含む方法。
２５．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が、約１５％～約２５％である、上記２４に記載の方法。
２６．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が、約２１％～約２５％である、上記２５に記載の方法。
２７．前記抗ＣＤ３８抗体のＧ０Ｆオリゴ糖含有量が、約６５％～約７４％である、上記２４に記載の方法。
２８．前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が、約６８％～７４％である、上記２５に記載の方法。
２９．前記培養培地におけるＭｎの濃度が、約４．０ｐｐｂ～約８．５ｐｐｂのＭｎを含むように調節される、上記２４に記載の方法。
３０．前記培養培地におけるＭｎの濃度が、約４．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調節される、上記２９に記載の方法。
３１．前記培養培地におけるＭｎの濃度が、約５．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調節される、上記３０に記載の方法。
３２．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約１５％～約２７％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６５％～約７４％であり、前記培養培地が、約４．０ｐｐｂ～約８．５ｐｐｂのＭｎを含むように調節される、上記２４～３０のいずれかに記載の方法。
３３．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約１５％～約２５％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６８％～約７４％であり、前記培養培地が、約４．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調節される、上記２４～３０のいずれかに記載の方法。
３４．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約２１％～約２５％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６８％～約７４％であり、前記培養培地が、約５．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調節される、上記２４～３０のいずれかに記載の方法。
３５．前記培養培地が、基本培地又は流加培地である、上記２４～３４のいずれかに記載の方法。
３６．培養が、流加培養又は灌流培養を含む、上記２４～３５のいずれかに記載の方法。
３７．前記宿主細胞が、真核細胞である、上記２４～３６のいずれかに記載の方法。
３８．前記真核細胞が、ＣＨＯ細胞、ＰＥＲ．Ｃ６細胞、ＮＳ０細胞、Ｓｐ２／０細胞、又はＢＨＫ細胞である、上記３７に記載の方法。
３９．前記ＣＨＯ細胞が、ＣＨＯ－Ｋ１細胞、ＣＨＯ－ＤＧ４４細胞、ＣＨＯ－Ｓ細胞、又はＣＨＯ－ＤＸＢ１１細胞である、上記３８に記載の方法。
４０．前記ＣＨＯ細胞が、ＧＳが不足している、上記３９に記載の方法。
４１．ＧＭＰに準拠する条件下で実施される、上記２４～４０のいずれかに記載の方法。
４２．前記抗ＣＤ３８抗体が、前記配列番号７のＶＨ及び前記配列番号９のＶＬを含む、上記２４～４１のいずれかに記載の方法。
４３．前記抗ＣＤ３８抗体が、ＩｇＧ１アイソタイプを含む、上記２４～４２のいずれかに記載の方法。
４４．前記抗ＣＤ３８抗体が、配列番号９のＨＣ及び配列番号１０のＬＣを含む、上記２４～４３のいずれかに記載の方法。
４５．前記抗ＣＤ３８抗体が、バイオ後続品である、上記２４～４４のいずれかに記載の方法。
４６．約１５％～約２７％のＧ１Ｆオリゴ糖含有量を有する、配列番号７のＶＨ及び配列番号８のＶＬをコードしているポリヌクレオチドから発現する抗ＣＤ３８抗体を含む医薬品の製造方法であって、
ａ）約８．５ｐｐｂ以下のＭｎを含む培養培地を調製することと、
ｂ）工程ａ）で調製された前記培養培地中で前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドを含む宿主細胞を培養することによって、前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量を制御し、それにより、前記約１５％～約２７％のＧ１Ｆオリゴ糖含有量を有する前記抗ＣＤ３８抗体を生成することと、
ｃ）前記抗ＣＤ３８抗体を医薬品として製剤化することと、
を含む方法。
４７．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が、約１５％～約２５％である、上記４６に記載の方法。
４８．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が、約２１％～約２５％である、上記４７に記載の方法。
４９．前記抗ＣＤ３８抗体のＧ０Ｆオリゴ糖含有量が、約６５％～約７４％である、上記４６～４９のいずれかに記載の方法。
５０．前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が、約６８％～７４％である、上記４６～４９のいずれかに記載の方法。
５１．前記培養培地を調製することが、前記培養培地を調製するために使用される原材料成分の１つ以上のバッチにおけるＭｎ濃度を測定することと、組み合わせて約８．５ｐｐｂ以下のＭｎを含有する原材料成分の１つ以上のバッチを選択することと、を含む、上記４６に記載の方法。
５２．前記培養培地が、約４．０ｐｐｂ～約８．５ｐｐｂのＭｎを含むように調製される、上記５１に記載の方法。
５３．前記培養培地が、約４．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記５２に記載の方法。
５４．前記培養培地が、約５．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記５３に記載の方法。
５５．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約１５％～約２７％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６５％～約７４％であり、前記培養培地が、約４．０ｐｐｂ～約８．５ｐｐｂのＭｎを含むように調製される、上記４６～５４のいずれかに記載の方法。
５６．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約１５％～約２５％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６８％～約７４％であり、前記培養培地が、約４．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記４６～５４のいずれかに記載の方法。
５７．前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量が約２１％～約２５％であり、前記抗ＣＤ３８抗体の前記Ｇ０Ｆオリゴ糖含有量が約６８％～約７４％であり、前記培養培地が、約５．０ｐｐｂ～約６．５ｐｐｂのＭｎを含むように調製される、上記４６～５４のいずれかに記載の方法。
５８．前記培養培地が、基本培地又は流加培地である、上記４６～５７のいずれかに記載の方法。
５９．培養が、流加培養又は灌流培養を含む、上記４６～５８のいずれかに記載の方法。
６０．前記宿主細胞が、真核細胞である、上記４６～５９のいずれかに記載の方法。
６１．前記真核細胞が、ＣＨＯ細胞、ＰＥＲ．Ｃ６細胞、ＮＳ０細胞、Ｓｐ２／０細胞、又はＢＨＫ細胞である、上記６０に記載の方法。
６２．前記ＣＨＯ細胞が、ＣＨＯ－Ｋ１細胞、ＣＨＯ－ＤＧ４４細胞、ＣＨＯ－Ｓ細胞、又はＣＨＯ－ＤＸＢ１１細胞である、上記６１に記載の方法。
６３．前記ＣＨＯ細胞が、グルタミン合成酵素（ＧＳ）が不足している、上記６２に記載の方法。
６４．ＧＭＰに準拠する条件下で実施される、上記４６～６３のいずれかに記載の方法。
６５．前記抗ＣＤ３８抗体が、前記配列番号７のＶＨ及び前記配列番号９のＶＬを含む、上記４６～６４のいずれかに記載の方法。
６６．前記抗ＣＤ３８抗体が、ＩｇＧ１アイソタイプを含む、上記４６～６５のいずれかに記載の方法。
６７．前記抗ＣＤ３８抗体が、配列番号９のＨＣ及び配列番号１０のＬＣを含む、上記４６～６６のいずれかに記載の方法。
６８．前記抗ＣＤ３８抗体が、バイオ後続品である、上記４６～６７のいずれかに記載の方法。
６９．前記医薬品を製剤化することが、ｐＨ約５．６で、約３０，０００Ｕ～約４５，０００Ｕの量の組み換えヒトヒアルロニダーゼ（ｒＨｕＰＨ２０）、約５ｍＭ～約１５ｍＭの濃度のヒスチジン、約１００ｍＭ～約３００ｍＭの濃度のソルビトール、約０．０１％ｗ／ｖ～約０．０４％ｗ／ｖの濃度のＰＳ－２０、及び約１ｍｇ／ｍＬ～約２ｍｇ／ｍＬの濃度のメチオニンを用いて、約２０ｍｇ／ｍＬ～約１８０ｍｇ／ｍＬの前記抗ＣＤ３８抗体を製剤化することを含む、上記４６～６８のいずれかに記載の方法。
７０．前記医薬品を製剤化することが、ｐＨ約５．６で、約２，０００Ｕ／ｍＬの組み換えヒトヒアルロニダーゼ（ｒＨｕＰＨ２０）、約５ｍＭ～約１５ｍＭのヒスチジン、約１００ｍＭ～約３００ｍＭのソルビトール、約０．０１％ｗ／ｖ～約０．０４％ｗ／ｖのＰＳ－２０、及び約１ｍｇ／ｍＬ～約２ｍｇ／ｍＬのメチオニン中で、約１２０ｍｇ／ｍＬの前記抗ＣＤ３８抗体を製剤化することを含む、上記４６～６９のいずれかに記載の方法。
７１．前記医薬品を製剤化することが、ｐＨ約５．５で、約２５ｍＭの酢酸、約６０ｍＭの塩化ナトリウム、約１４０ｍＭのマンニトール、及び約０．０４％ｗ／ｖのポリソルベート－２０（ＰＳ－２０）中で、２０ｍｇ／ｍＬの前記ＣＤ３８抗体を製剤化することを含む、上記４６～６８のいずれかに記載の方法。
While the disclosure has been described in general terms, embodiments of the disclosure are further disclosed in the following examples, which should not be construed as limiting the scope of the claims.
Various embodiments of the invention are presented below.
1. Antibodies expressed from polynucleotides encoding the heavy chain variable region (VH) of SEQ ID NO:7 and the light chain variable region (VL) of SEQ ID NO:8 having a G1F oligosaccharide content of about 15% to about 27%. A method of producing a CD38 antibody, comprising:
a) preparing a culture medium comprising about 8.5 parts per billion (ppb) or less of manganese (Mn);
b) production of said anti-CD38 antibody by culturing host cells comprising said polynucleotides encoding said VH of SEQ ID NO:7 and said VL of SEQ ID NO:8 in said culture medium prepared in step a); said poly encoding said VH of SEQ ID NO: 7 and said VL of SEQ ID NO: 8 that controls said G1F oligosaccharide content, thereby having a G1F oligosaccharide content of from about 15% to about 27%; generating said anti-CD38 antibody expressed from nucleotides;
method including.
2. 2. The method of claim 1, wherein said G1F oligosaccharide content of said anti-CD38 antibody is from about 15% to about 25%.
3. 2. The method of claim 1, wherein said G1F oligosaccharide content of said anti-CD38 antibody is from about 21% to about 25%.
4. 4. The method of any one of 1-3 above, wherein the anti-CD38 antibody has a G0F oligosaccharide content of about 65% to about 74%.
5. 4. The method of any of claims 1-3, wherein said G0F oligosaccharide content of said anti-CD38 antibody is about 68%-74%.
6. preparing the culture medium comprises measuring the Mn concentration in one or more batches of raw material components used to prepare the culture medium, and a raw material containing less than or equal to about 8.5 ppb Mn and selecting one or more batches of ingredients.
7. 7. The method of claim 6, wherein said culture medium is prepared to contain Mn from about 4.0 ppb to about 8.5 ppb.
8. 8. The method of claim 7, wherein the culture medium is prepared to contain Mn from about 4.0 ppb to about 6.5 ppb.
9. 9. The method of claim 8, wherein said culture medium is prepared to contain Mn from about 5.0 ppb to about 6.5 ppb.
10. The G1F oligosaccharide content of the anti-CD38 antibody is about 15% to about 27%, the G0F oligosaccharide content of the anti-CD38 antibody is about 65% to about 74%, and the culture medium contains about 10. The method of any one of 1-9 above, prepared to contain from 4.0 ppb to about 8.5 ppb of Mn.
11. The G1F oligosaccharide content of the anti-CD38 antibody is about 15% to about 25%, the G0F oligosaccharide content of the anti-CD38 antibody is about 68% to about 74%, and the culture medium contains about 10. The method of any one of 1-9 above, prepared to contain from 4.0 ppb to about 6.5 ppb of Mn.
12. The G1F oligosaccharide content of the anti-CD38 antibody is about 21% to about 25%, the G0F oligosaccharide content of the anti-CD38 antibody is about 68% to about 74%, and the culture medium contains about 10. A method according to any one of 1 to 9 above, prepared to contain from 5.0 ppb to about 6.5 ppb of Mn.
13. 13. The method according to any one of 1 to 12 above, wherein the culture medium is a basal medium or a feed medium.
14. 14. The method according to any one of 1 to 13 above, wherein the culture comprises fed-batch culture or perfusion culture.
15. 15. The method according to any one of 1 to 14 above, wherein the host cell is a eukaryotic cell.
16. Said eukaryotic cells are CHO cells, PER. 17. The method of 16 above, wherein the cells are C6 cells, NS0 cells, Sp2/0 cells, or BHK cells.
17. 17. The method of 16 above, wherein the CHO cells are CHO-K1 cells, CHO-DG44 cells, CHO-S cells, or CHO-DXB11 cells.
18. 18. The method of Claim 17, wherein said CHO cells are deficient in glutamine synthetase (GS).
19. 19. The method according to any one of 1 to 18 above, which is carried out under GMP-compliant conditions.
20. 20. The method of any one of 1 to 19 above, wherein said anti-CD38 antibody comprises said VH of SEQ ID NO:7 and said VL of SEQ ID NO:9.
21. 21. The method of any of 1-20 above, wherein said anti-CD38 antibody comprises an IgG1 isotype.
22. 22. The method of any one of 1-21 above, wherein said anti-CD38 antibody comprises a heavy chain (HC) of SEQ ID NO:9 and a light chain (LC) of SEQ ID NO:10.
23. 23. The method of any of 1-22 above, wherein said anti-CD38 antibody is a biosimilar.
24. A method of generating an anti-CD38 antibody expressed from a polynucleotide encoding the VH of SEQ ID NO:7 and the VL of SEQ ID NO:8 having a G1F oligosaccharide content of about 15% to about 27%, comprising:
a) culturing host cells expressing said polynucleotides encoding said VH of SEQ ID NO:7 and said VL of SEQ ID NO:8 under conditions in which said anti-CD38 antibody is generated;
b) the G1F of said anti-CD38 antibody by monitoring the concentration of Mn in the culture medium during biosynthesis of said anti-CD38 antibody and adjusting the concentration of Mn in said culture medium during biosynthesis of said anti-CD38 antibody; controlling the oligosaccharide content and adjusting the concentration of Mn in the culture medium to contain about 8.5 ppb or less of Mn, thereby having a G1F oligosaccharide content of about 15% to about 27%; producing an anti-CD38 antibody;
method including.
25. 25. The method of Claim 24, wherein said G1F oligosaccharide content of said anti-CD38 antibody is from about 15% to about 25%.
26. 26. The method of Claim 25, wherein said G1F oligosaccharide content of said anti-CD38 antibody is from about 21% to about 25%.
27. 25. The method of 24 above, wherein the G0F oligosaccharide content of said anti-CD38 antibody is from about 65% to about 74%.
28. 26. The method of Claim 25, wherein said G0F oligosaccharide content of said anti-CD38 antibody is between about 68% and 74%.
29. 25. The method of Claim 24, wherein the concentration of Mn in said culture medium is adjusted to contain from about 4.0 ppb to about 8.5 ppb of Mn.
30. 30. The method of Claim 29, wherein the concentration of Mn in said culture medium is adjusted to contain from about 4.0 ppb to about 6.5 ppb of Mn.
31. 31. The method of Claim 30, wherein the concentration of Mn in said culture medium is adjusted to contain from about 5.0 ppb to about 6.5 ppb of Mn.
32. The G1F oligosaccharide content of the anti-CD38 antibody is about 15% to about 27%, the G0F oligosaccharide content of the anti-CD38 antibody is about 65% to about 74%, and the culture medium contains about 31. The method of any of 24-30, wherein the method is adjusted to contain Mn from 4.0 ppb to about 8.5 ppb.
33. The G1F oligosaccharide content of the anti-CD38 antibody is about 15% to about 25%, the G0F oligosaccharide content of the anti-CD38 antibody is about 68% to about 74%, and the culture medium contains about 31. The method of any of 24-30 above, wherein the method is adjusted to contain Mn from 4.0 ppb to about 6.5 ppb.
34. The G1F oligosaccharide content of the anti-CD38 antibody is about 21% to about 25%, the G0F oligosaccharide content of the anti-CD38 antibody is about 68% to about 74%, and the culture medium contains about 31. The method of any of 24-30, wherein the method is adjusted to contain Mn from 5.0 ppb to about 6.5 ppb.
35. 35. The method according to any one of 24 to 34 above, wherein the culture medium is a basal medium or a feed medium.
36. 36. The method according to any of the above 24-35, wherein the culture comprises fed-batch culture or perfusion culture.
37. 37. The method of any of 24-36 above, wherein said host cell is a eukaryotic cell.
38. Said eukaryotic cells are CHO cells, PER. 38. The method of 37 above, wherein the cells are C6 cells, NS0 cells, Sp2/0 cells, or BHK cells.
39. 39. The method of 38 above, wherein the CHO cells are CHO-K1 cells, CHO-DG44 cells, CHO-S cells, or CHO-DXB11 cells.
40. 40. The method of 39 above, wherein said CHO cells are GS deficient.
41. 41. The method of any of 24-40 above, which is carried out under GMP-compliant conditions.
42. 42. The method of any of 24-41 above, wherein said anti-CD38 antibody comprises said VH of SEQ ID NO:7 and said VL of SEQ ID NO:9.
43. 43. The method of any of 24-42 above, wherein said anti-CD38 antibody comprises an IgG1 isotype.
44. 44. The method of any of 24-43 above, wherein said anti-CD38 antibody comprises HC of SEQ ID NO:9 and LC of SEQ ID NO:10.
45. 45. The method of any of 24-44 above, wherein said anti-CD38 antibody is a biosimilar.
46. A method for producing a medicament comprising an anti-CD38 antibody expressed from a polynucleotide encoding VH of SEQ ID NO:7 and VL of SEQ ID NO:8 having a G1F oligosaccharide content of about 15% to about 27%, comprising:
a) preparing a culture medium comprising about 8.5 ppb or less of Mn;
b) production of said anti-CD38 antibody by culturing host cells comprising said polynucleotides encoding said VH of SEQ ID NO:7 and said VL of SEQ ID NO:8 in said culture medium prepared in step a); controlling said G1F oligosaccharide content, thereby producing said anti-CD38 antibody having a G1F oligosaccharide content of said about 15% to about 27%;
c) formulating said anti-CD38 antibody as a pharmaceutical;
method including.
47. 47. The method of 46, wherein said G1F oligosaccharide content of said anti-CD38 antibody is from about 15% to about 25%.
48. 48. The method of 47 above, wherein said G1F oligosaccharide content of said anti-CD38 antibody is from about 21% to about 25%.
49. 49. The method of any of 46-49 above, wherein the anti-CD38 antibody has a G0F oligosaccharide content of about 65% to about 74%.
50. 49. The method of any of 46-49, wherein the G0F oligosaccharide content of the anti-CD38 antibody is about 68%-74%.
51. preparing the culture medium comprises measuring the Mn concentration in one or more batches of raw material components used to prepare the culture medium, and a raw material containing less than or equal to about 8.5 ppb Mn 47. The method of claim 46, comprising selecting one or more batches of ingredients.
52. 52. The method of Claim 51, wherein said culture medium is prepared to contain from about 4.0 ppb to about 8.5 ppb Mn.
53. 53. The method of claim 52, wherein said culture medium is prepared to contain Mn from about 4.0 ppb to about 6.5 ppb.
54. 54. The method of claim 53, wherein said culture medium is prepared to contain from about 5.0 ppb to about 6.5 ppb Mn.
55. The G1F oligosaccharide content of the anti-CD38 antibody is about 15% to about 27%, the G0F oligosaccharide content of the anti-CD38 antibody is about 65% to about 74%, and the culture medium contains about 55. The method of any of 46-54 above, prepared to contain from 4.0 ppb to about 8.5 ppb of Mn.
56. The G1F oligosaccharide content of the anti-CD38 antibody is about 15% to about 25%, the G0F oligosaccharide content of the anti-CD38 antibody is about 68% to about 74%, and the culture medium contains about 55. The method of any of 46-54 above, prepared to contain from 4.0 ppb to about 6.5 ppb of Mn.
57. The G1F oligosaccharide content of the anti-CD38 antibody is about 21% to about 25%, the G0F oligosaccharide content of the anti-CD38 antibody is about 68% to about 74%, and the culture medium contains about 55. The method of any of 46-54 above, prepared to contain from 5.0 ppb to about 6.5 ppb of Mn.
58. 58. The method of any of 46-57 above, wherein the culture medium is a basal medium or a feed medium.
59. 59. The method of any of 46-58 above, wherein the culture comprises fed-batch culture or perfusion culture.
60. 60. The method of any of 46-59 above, wherein said host cell is a eukaryotic cell.
61. Said eukaryotic cells are CHO cells, PER. 61. The method of 60 above, wherein the cells are C6 cells, NS0 cells, Sp2/0 cells, or BHK cells.
62. 62. The method of 61 above, wherein the CHO cells are CHO-K1 cells, CHO-DG44 cells, CHO-S cells, or CHO-DXB11 cells.
63. 63. The method of 62 above, wherein said CHO cells are deficient in glutamine synthetase (GS).
64. 64. The method of any of 46-63 above, which is carried out under GMP-compliant conditions.
65. 65. The method of any of 46-64 above, wherein said anti-CD38 antibody comprises said VH of SEQ ID NO:7 and said VL of SEQ ID NO:9.
66. 66. The method of any of 46-65 above, wherein said anti-CD38 antibody comprises an IgG1 isotype.
67. 67. The method of any of 46-66 above, wherein said anti-CD38 antibody comprises HC of SEQ ID NO:9 and LC of SEQ ID NO:10.
68. 68. The method of any of 46-67 above, wherein said anti-CD38 antibody is a biosimilar.
69. Formulating the pharmaceutical comprises recombinant human hyaluronidase (rHuPH20) in an amount of about 30,000 U to about 45,000 U, histidine in a concentration of about 5 mM to about 15 mM, about 100 mM to about 300 mM, at a pH of about 5.6. sorbitol at a concentration of about 0.01% w/v to about 0.04% w/v, PS-20 at a concentration of about 0.01% w/v to about 0.04% w/v, and methionine at a concentration of about 1 mg/mL to about 2 mg/mL. 69. The method of any of 46-68 above, comprising formulating from mL to about 180 mg/mL of said anti-CD38 antibody.
70. Formulating the pharmaceutical comprises about 2,000 U/mL recombinant human hyaluronidase (rHuPH20), about 5 mM to about 15 mM histidine, about 100 mM to about 300 mM sorbitol, about 0.01 at pH about 5.6. formulating about 120 mg/mL of said anti-CD38 antibody in % w/v to about 0.04% w/v PS-20 and about 1 mg/mL to about 2 mg/mL methionine; The method according to any one of 46 to 69 above.
71. Formulating the pharmaceutical comprises about 25 mM acetic acid, about 60 mM sodium chloride, about 140 mM mannitol, and about 0.04% w/v polysorbate-20 (PS-20) at a pH of about 5.5. 69. The method of any of 46-68 above, comprising formulating 20 mg/mL of said CD38 antibody therein.
Claims (39)
ａ）２十億分率（ｐｐｂ）～８．５ｐｐｂのマンガン（Ｍｎ）を含む培養培地を調製することと、
ｂ）工程ａ）で調製された前記培養培地中で前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドを含む宿主細胞を培養することによって、前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量を制御し、それにより、前記１５％～２７％のＧ１Ｆオリゴ糖含有量を有する、前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドから発現する前記抗ＣＤ３８抗体を生成することと、
を含む方法。 The heavy chain variable region (VH ) of SEQ ID NO: 7 and the light chain variable region of SEQ ID NO:8 ( A method for generating an anti-CD38 antibody expressed from a polynucleotide encoding VL), comprising:
a) 2 parts per billion (ppb) to 8. preparing a culture medium comprising 5ppb manganese (Mn);
b) production of said anti-CD38 antibody by culturing host cells comprising said polynucleotides encoding said VH of SEQ ID NO:7 and said VL of SEQ ID NO:8 in said culture medium prepared in step a); said encoding the VH of SEQ ID NO: 7 and the VL of SEQ ID NO: 8 controlling said G1F oligosaccharide content, thereby having a G1F oligosaccharide content of said 15% to 27%; producing said anti-CD38 antibody expressed from a polynucleotide;
method including.
ａ）請求項１に記載の方法にしたがって抗ＣＤ３８抗体を生成することと、
ｂ）前記抗ＣＤ３８抗体を医薬品として製剤化することと、
を含む方法。 A method for manufacturing a pharmaceutical,
a) generating an anti-CD38 antibody according to the method of claim 1;
b) formulating said anti-CD38 antibody as a medicament;
method including.
ａ）前記抗ＣＤ３８抗体が生成される条件下で前記配列番号７のＶＨ及び前記配列番号８のＶＬをコードしている前記ポリヌクレオチドを発現する宿主細胞を培養することと、
ｂ）前記抗ＣＤ３８抗体の生合成中の培養培地におけるＭｎの濃度をモニタリングし、前記抗ＣＤ３８抗体の生合成中の前記培養培地におけるＭｎの濃度を調節することによって、前記抗ＣＤ３８抗体の前記Ｇ１Ｆオリゴ糖含有量を制御し、前記培養培地におけるＭｎの濃度を２ｐｐｂ～８．５ｐｐｂ以下のＭｎを含むように調節し、それにより、前記１５％～２７％のＧ１Ｆオリゴ糖含有量を有する前記抗ＣＤ３８抗体を生成することと、
を含む方法。 A method for generating an anti-CD38 antibody expressed from a polynucleotide encoding VH of SEQ ID NO:7 and VL of SEQ ID NO:8 having a G1F oligosaccharide content of 15 % to 27%, comprising:
a) culturing host cells expressing said polynucleotides encoding said VH of SEQ ID NO:7 and said VL of SEQ ID NO:8 under conditions in which said anti-CD38 antibody is generated;
b) the G1F of said anti-CD38 antibody by monitoring the concentration of Mn in the culture medium during biosynthesis of said anti-CD38 antibody and adjusting the concentration of Mn in said culture medium during biosynthesis of said anti-CD38 antibody; The oligosaccharide content is controlled and the concentration of Mn in said culture medium is adjusted to contain no more than 2 ppb to 8.5 ppb Mn, thereby reducing said 15% to 27% G1F oligosaccharide content. generating said anti-CD38 antibody having
method including.
原材料成分の１つ以上のバッチにおけるＭｎ濃度を測定することと、
組み合わせて２ｐｐｂ～８．５ｐｐｂのＭｎを含有する原材料成分の１つ以上のバッチを選択することと、
選択された前記原材料成分の１つ以上のバッチを、培養培地を調製するために使用することと、
を含む、請求項１に記載の方法。 preparing the culture medium,
measuring the Mn concentration in one or more batches of raw material ingredients;
selecting one or more batches of raw material ingredients that in combination contain between 2 ppb and 8.5 ppb of Mn;
using one or more batches of the selected raw material components to prepare a culture medium;
2. The method of claim 1 , comprising:
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2019
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