JP5773352B2 - 抗muc1抗体 - Google Patents
抗muc1抗体 Download PDFInfo
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- JP5773352B2 JP5773352B2 JP2010535828A JP2010535828A JP5773352B2 JP 5773352 B2 JP5773352 B2 JP 5773352B2 JP 2010535828 A JP2010535828 A JP 2010535828A JP 2010535828 A JP2010535828 A JP 2010535828A JP 5773352 B2 JP5773352 B2 JP 5773352B2
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- antibody
- pro
- antigen
- muc1
- ala
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- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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Description
A)2,3ST糖ペプチド(HGVTSAPDTRPAPGSTAPPA(配列番号1)のアミノ酸配列に従ったエピトープ)を提供する工程;
B)該2,3ST糖ペプチドで動物を免疫して、ハイブリドーマを得る工程;および
C)該動物から2,3ST(糖)に親和性を示すハイブリドーマのクローンを選択する工程;を包含する、方法を提供する。
本明細書において「タンパク質」、「ポリペプチド」、「オリゴペプチド」および「ペプチド」は、本明細書において同じ意味で使用され、任意の長さのアミノ酸のポリマーをいう。このポリマーは、直鎖であっても分岐していてもよく、環状であってもよい。アミノ酸は、天然のものであっても非天然のものであってもよく、改変されたアミノ酸であってもよい。この用語はまた、複数のポリペプチド鎖の複合体へとアセンブルされたものを包含し得る。この用語はまた、天然または人工的に改変されたアミノ酸ポリマーも包含する。そのような改変としては、例えば、ジスルフィド結合形成、グリコシル化、脂質化、アセチル化、リン酸化または任意の他の操作もしくは改変(例えば、標識成分との結合体化)。この定義にはまた、例えば、アミノ酸の1または2以上のアナログを含むポリペプチド(例えば、非天然のアミノ酸などを含む)、ペプチド様化合物(例えば、ペプトイド)および当該分野において公知の他の改変が包含される。
2,3ST6L:Neu5Acα2→3Galβ1→3[Galβ1→4GlcNAcβ1→6]GalNAcα-R2,3ST6SL:Neu5Acα2→3Galβ1→3[Neu5Acα2→3Galβ1→4GlcNAcβ1→6]GalNAcα-R
2,3ST:Neu5Acα2→3Galβ1→3GalNAcα-R
Tn:GalNAcα-R
T:Galβ1→3GalNAcα-R
本明細書において遺伝子の「相同性」とは、2以上の遺伝子配列の、互いに対する同一性の程度をいう。従って、ある2つの遺伝子の相同性が高いほど、それらの配列の同一性または類似性は高い。2種類の遺伝子が相同性を有するか否かは、配列の直接の比較、または核酸の場合ストリンジェントな条件下でのハイブリダイゼーション法によって調べられ得る。2つの遺伝子配列を直接比較する場合、その遺伝子配列間でDNA配列が、代表的には少なくとも80%同一である場合、好ましくは少なくとも90%同一である場合、より好ましくは少なくとも95%同一である場合、それらの遺伝子は相同性を有する。
A:G,I,V,L
C:M(含Sアミノ酸)
D:N、QまたはE
E:N、QまたはD
F:Y、Aなど
G:A
H:Wなど
I:A,L,V,(G)
K:R
L:A,I,V,(G)
M:Sなど
N:E、DまたはQ
P:HyP
Q:N、EまたはD
R:K
S:T、Y
T:S,Y
V:I,L,A、(G)
W:H
Y:F、S,T
これらのアミノ酸の間の置換は、本明細書において「保存的置換」ともいう。
本明細書において「抗体」とは、免疫反応において、抗原の刺激によって生体内でつくられ、抗原と特異的に結合したり反応したりするタンパク質またはそれらと同じ配列を有するものを化学合成等で生産したものを総称していう。実体は免疫グロブリンであり、Abとも称する。
ルガラクトサミンであり、Rは非糖部分である。
本明細書では、格別に他の意味であると指示しない限り、抗体などの任意のポリペプチド鎖は、N終末端(N−terminal extremity)で開始しC終末端(C−terminal extremity)で終結するアミノ酸配列を有するものとして記載する。抗原結合部位がVHおよびVLドメイン両方を含むとき、これらは同一ポリペプチド分子に位置し得るか、好ましくは、各ドメインは別の鎖に位置し得、この場合、VHドメインは免疫グロブリンすなわち抗体の重鎖またはその断片の一部であり、またVLは免疫グロブリンすなわち抗体の軽鎖またはその断片の一部である。
本発明の抗体は、当該分野において周知の任意の方法を用いて生産することができる。そのような方法の例示は、実施例に記載されるがそれに限定されない。まず、抗原を用いて動物を免疫することによって、抗体が産生される。
本免疫測定方法に用いる単一特異的な抗体としては、安定的に供給しうるモノクローナル抗体が望ましいがそれに限定されず、任意の分子を用いることができる。以下、モノクローナル抗体を用いて例示する。抗体(第1のモノクローナル抗体)を固相に固定し、抗原を含む資料と共にインキュベートする工程、さらに標識した第2のモノクローナル抗体を加えて、得られた混合物をインキュベートする工程、および混合物中の生成した標識された抗原抗体複合体を検出する工程を包含するサンドイッチ免疫学的測定法が例示される。また、本発明の免疫学的測定法では、試料と、固相化した第1のモノクローナル抗体および標識した第2のモノクローナル抗体とを同時にインキュベートしてもよい。サンドイッチ免疫学的測定法としては、その検出方法により、サンドイッチ放射免疫測定法(RIA法)、サンドイッチ酵素免疫測定法(EIA法)、サンドイッチ蛍光免疫測定法(FIA法)、サンドイッチ発光免疫測定法(CLIA法)、サンドイッチ発光酵素免疫測定法(CLEIA法)、サンドイッチ法に基づく免疫クロマトグラフ法などの全てのサンドイッチ免疫測定法が応用しうる。定量のためには、RIA法、EIA法が好ましい。
細胞傷害活性(cytotoxicity,%)=(実験値−自然遊離)/(最大遊離−自然遊離)×100
当該分野での技術水準によれば、当業者は、例えばCDRグラフティング法(例えば、EP 239400)により、ヒト化抗体を作製することが出来る。
本発明の化合物またはその製薬上許容される塩は、そのまま単独で投与することも可能であるが、通常各種の医薬製剤として提供するのが好ましい。また、それら医薬製剤は、動物および人に使用される。
1つの局面において、本発明は、本発明の抗体、その抗原結合性断片またはMUC1結合分子またはMUC1結合分子を用いるがんの診断法、本発明の抗体、その抗原結合性断片またはMUC1結合分子またはMUC1結合分子を含む診断剤、あるいは抗体、その抗原結合性断片またはMUC1結合分子またはMUC1結合分子を含む診断キットを提供する。本発明のがんの診断法、診断剤または診断キットにおいて含まれる抗体、その抗原結合性断片またはMUC1結合分子またはMUC1結合分子は、上述した本発明の抗体その抗原結合性断片またはMUC1結合分子またはMUC1結合分子の任意の実施形態でありうることが理解される。本発明の抗体、抗原結合性断片またはMUC1結合分子またはMUC1結合分子は、特定のがんに特異的に結合することから、これらのがんの診断に使用されうる。
DMF:N,N-ジメチルホルムアミド
DCM:ジクロロメタン
HBTU:1-[ビス(ジメチルアミノ)メチレン]-1H-ベンゾトリアゾリウム-3-オキシドヘキサフルオロホスファート
HOBt:N-ヒドロキシベンゾトリアゾール
DIEA:ジイソプロピルエチルアミン
Fmoc:(9H-フルオレン-9-イル)メトキシカルボニル
TIS:トリイソプロピルシラン
CMP-NANA:シチジン-5’-モノホスホ-N-アセチルノイラミン酸2ナトリウム
UDP-Gal:ウリジン-5’-ジホスホ-N-ガラクトース2ナトリウム
本実施例のマイクロ波照射下における反応は、マイクロ波式有機化学合成装置グリーン・モチーフ・I(東京電子株式会社製)を用いた。
実施例化合物1の合成
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Galβ1→3GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(1)
糖ペプチド固相合成は、固相担体としてRink Amide−PEGA樹脂(0.05mmol/g,500mg,25μmol)を用いた。アミノ酸伸長反応は、マイクロ波照射(40W、2450MHz、50℃)の条件下、Fmocアミノ酸誘導体(75μmol)、HBTU(75μmol)とHOBt(75μmol)、DIEA(150μmol)のDMF溶液で5分間反応した。糖鎖置換アミノ酸伸長反応は、Fmoc−Thr(Ac6 core1)−OH:N−α−Fmoc−O−[O−(2,3,4,6−テトラ−O−アセチル−β−D−ガラクトピラノシル)−(1→3)]−4,6−ジ−O−アセチル−2−アセトアミド−2−デオキシ−α−D−ガラクトピラノシル}−L−スレオニンを1.5等量使用して同様の条件下20分間反応した。13mM
HOBtの無水酢酸/DIEA/DMF(4.75:2.25:93 v/v/v)溶液で室温下5分間処理して未反応のアミノ基のアセチル化を実施した。続いて、マイクロ波照射(40W、2450MHz、50℃)の条件下、20%ピペリジン/DMFで3分間処理してFmoc基を脱保護した。糖ペプチドの合成は、これら3工程(1)各種Fmocアミノ酸での伸長、(2)アセチル化処理、(3)脱Fmoc化を順次繰り返した。得られた固相樹脂をトリフルオロ酢酸:水:TIS(93:5:2 v/v/v)で1時間処理した。反応液を濾過し溶媒を留去した後、得られた残渣にエーテルを加えて沈殿させて、粗結晶を得た。粗生成物を逆相高速液体クロマトグラフィーにより精製を行い、糖のアセチル保護体を得た。得られた保護体をメタノールに溶解して、1N 水酸化ナトリウム水溶液を加えてpH 12.0−12.5に調整して室温下1時間処理した。10%酢酸を加えてpH7付近に調整した後、溶媒を留去した。得られた残渣を逆相高速液体クロマトグラフィーにより精製を行い、凍結乾燥粉末として化合物1を得た。
凍結乾燥粉末 (26 mg, 収率 46 %). MALDI-TOF MS: m/zcalcd for C94H152N27O37[M+H]+2251.1, found 2250.7. ESI-HRMS: m/z calcd for C94H149N27O37[M-2H]2-1124.0304, found 1124.0325 [M-2H]2-. Amino acid analysis:Ala(4) 3.9,Asp(1) 1.0, Arg(1) 1.0, Gly(2) 2.0, His(1) 1.1, Pro(5) 5.4, Ser(2)1.7, Thr(3)2.8, Val(1) 1.0.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Neu5Acα2→3Galβ1→3GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(2)
化合物1(22.5mg)、CMP−NANA(32.9mg)とα2,3−(O)−シアル酸糖転移酵素(44mU)の50mMHEPES緩衝液(10mM MnCl2,0.1%BSA,pH7.0)(5.0ml)溶液の条件下、25℃で24時間静置反応した。反応混合物は、逆相高速液体クロマトグラフィーにより精製して凍結乾燥粉末として化合物2を得た。
凍結乾燥粉末 (20 mg, 収率 80 %). MALDI-TOF MS: m/zcalcd for C94H152N27O37[M+H]+2251.1, found 2250.7. ESI-HRMS: m/z calcd for C94H149N27O37[M-2H]2-1124.0304, found 1124.0325 [M-2H]2-. Amino acid analysis:Ala(4) 3.9,Asp(1) 1.0, Arg(1) 1.0, Gly(2) 2.0, His(1) 1.1, Pro(5) 5.4, Ser(2)1.7, Thr(3)2.8, Val(1) 1.0.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(3)
化合物1と同様にして、Fmoc−Thr(Ac3Tn)−OH:N−α−Fmoc−O−(3,4,6−トリ−O−アセチル−2−アセトアミド−2−デオキシ−α−D−ガラクトピラノシル)−L−スレオニンを用い合成した。
凍結乾燥粉末 (15 mg, 収率 14 %). MALDI-TOFMS: m/z calcd for C88H141N27O32[M+H]+2089.0, found 2089.1. ESI-HRMS: m/z calcdfor C88H143N27O32[M+3H]3+697.0157, found 697.0174. Aminoacid analysis: Ala(4) 4.0, Asp(1) 1.0,Arg(1) 1.0, Gly(2) 1.9, His(1) 1.0,Pro(5) 5.2, Ser(2) 1.7, Thr(3) 2.8, Val(1)1.0.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Neu5Acα2→6GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(4)
化合物1と同様にして、Fmoc−Thr(Ac6 Sialyl Tn)−OH:N−α−Fmoc−O−{[メチル−(5−アセトアミド−4,7,8,9−テトラ−O−アセチル−3,5−ジデオキシ−α−D−グリセロ−D−ガラクト−2−ノヌロピラノシル)オナト−(2→6)]−3,4−ジ−O−アセチル−2−アセトアミド−2−デオキシ−α−D−ガラクトピラノシル}−L−スレオニンを用いて、合成した。糖部分の脱アセチル化を行った後、水に溶解して1N 水酸化ナトリウム水溶液を加えてpH 12.0以下に調整して室温下6時間処理した。10%酢酸を加えてpH7付近に調整した後、精製を行い凍結乾燥粉末として化合物4を得た。
凍結乾燥粉末(2 mg, 収率15 %). MALDI-TOF MS: m/z calcd for C99H158N28O40[M+H]+2380.1, found 2380.1. ESI-HRMS: m/z calcd for C99H158N28O40[M+3H]3+794.0475, found 794.0494. Amino acid analysis: Ala(4) 3.9, Asp(1)1.0, Arg(1) 1.0, Gly(2) 1.9,His(1) 0.8, Pro(5) 5.1, Ser(2) 1.7, Thr(3) 2.8,Val(1) 1.0.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Galβ1→3[GlcNAcβ1→6]GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(5)
化合物1と同様にして、Fmoc−Thr(Ac7 core2−OH:N−α−Fmoc−O−{(2,3,4,6−テトラ−O−アセチル−β−D−ガラクトピラノシル)−(1→3)−O−[2−アセトアミド−3,4,6−トリ−O−アセチル−2−デオキシ−β−D−グルコピラノシル−(1→6)]−2−アセトアミド−2−デオキシ−α−D−ガラクトピラノシル}−L−スレオニンを用いて化合物5を得た。
凍結乾燥粉末 (51 mg, 収率: 52 %). MALDI-TOFMS: m/z calcd for C102H165N28O42[M+H]+2454.2, found 2454.5. ESI-HRMS: m/z calcd for C102H163N28O42[M-H]-2452.1479, found 2452.1475. Amino acid analysis: Ala(4) 3.7, Asp(1)1.0, Arg(1)1.0, Gly(2) 1.9, His(1) 0.9, Pro(5) 5.2, Ser(2) 1.6, Thr(3) 2.6,Val(1) 0.9.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Neu5Acα2→3Galβ1→3[GlcNAcβ1→6]GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(6)
化合物5およびCMP−NANAとα2,3−(O)−シアル酸転移酵素(5mU/ml)の50mM HEPES buffer(10mM MnCl2,0.1%BSA,pH7.0)溶液で反応して化合物6を得た。
凍結乾燥粉末(6 mg, quant.), MALDI-TOFMS: m/z calcd for C113H182N29O50:[M+H]+2745.3,found 2745.8. ESI-HRMS: m/z calcd for C113H180N29O50[M-H]-2743.2434, found 2743.2410.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Neu5Acα2→3Galβ1→3[Galβ1→4GlcNAcβ1→6]GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(7)
化合物5およびUDP−Gal,CMP−NANA、β1,4−ガラクトース転移酵素(100mU/ml)とα2,3−(O)−シアル酸転移酵素(5mU/ml)の50mM HEPES buffer(10mM MnCl2,0.1%BSA,pH7.0)溶液で反応して化合物7を得た。
凍結乾燥粉末(7 mg, quant.), MALDI-TOF MS: m/z calcd for C119H192N29O55[M+H]+2907.3, found 2906.5. ESI-HRMS: m/z calcd for C119H190N29O55[M-H]-2905.2962, found 2905.2922.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Neu5Acα2→3Galβ1→3[Neu5Acα2→3Galβ1→4GlcNAcβ1→6]GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(8)
化合物5およびUDP−Gal,CMP−NANA、β1,4−ガラクトース転移酵素(100mU/ml)、α2,3−(O)−シアル酸転移酵素(5mU/ml)とα2,3−(N)−シアル酸転移酵素(74mU/ml)の50mM HEPES buffer(10mM MnCl2,0.1%BSA,pH7.0)溶液で反応して化合物8を得た。
凍結乾燥粉末(7 mg, 収率46 %). MALDI-TOF MS: m/z calcd for C130H209N30O63[M+H]+3198.4, found 3198.0. ESI-HRMS: m/z calcd for C130H207N30O63[M-H]-3196.3916, found 3196.3899.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GlcNAcβ1→6GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(9)
化合物4と同様の条件下、Fmoc−Thr(Ac5 core6)−OH:N−α−Fmoc−O−{[3,4,6−トリ−O−アセチ−2−アセトアミド−2−デオキシ−β−D−グルコピラノシル−(1→6)]−3,4−ジ−O−アセチル−2−アセトアミド−2−デオキシ−α−D−ガラクトピラノシル}−L−スレオニンを用いて合成した。凍結乾燥粉末(17 mg, 収率 30 %). MALDI-TOF MS: m/zcalcd for C96H155N28O37[M+H]+2292.1, found 2290.6. ESI-HRMS: m/z calcd for C96H154N28O37[M+2H]2+1146.5593, found 1146.5568. Amino acid analysis: Ala(4) 4.1, Asp(1)1.0, Arg(1) 1.0, Gly(2) 2.0,His(1) 1.0, Pro(5) 5.4, Ser(2) 1.7, Thr(3) 2.8,Val(1) 1.0.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Galβ1→3[NeuAcα2→6]GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(10)
化合物1と同様にして、Fmoc−Thr(Ac6 2,6−Sialyl T)−OH:N−α−Fmoc−O−{[(2,3,4,6−テトラ−O−アセチル−β−D−ガラクトピラノシル)−(1→3)]−O−[メチル−(5−アセトアミド−4,7,8,9−テトラ−O−アセチル−3,5−ジデオキシ−α−D−グリセロ−D−ガラクト−2−ノヌロピラノシル)オナト−(2→6)]−2−アセトアミド−2−デオキシ−α−D−ガラクトピラノシル}−L−スレオニンを用いて化合物8を得た。
凍結乾燥粉末 (6 mg, 収率 39 %). MALDI-TOF MS: m/zcalcd for C105H169N28O45[M+H]+2542.2, found 2452.6. ESI-HRMS: m/z calcd for C105H170N28O45[M+2H]2+1271.5937, found 1271.5945. Amino acid analysis:Ala(4) 3.8, Asp(1) 1.0,Arg(1) 1.0, Gly(2) 1.9, His(1) 0.9, Pro(5) 5.2, Ser(2)1.6, Thr(3) 2.7, Val(1)0.9.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(NeuAcα2→3Galβ1→3[NeuAcα2→6]GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(11)
化合物2と同様の条件により合成した。
凍結乾燥粉末(2 mg, quant.). MALDI-TOFMS: m/z calcd for C116H186N29O53[M+H]+2833.3, found 2833.2. ESI-HRMS:m/z calcd for C116H1187N29O53[M+2H]2+1417.1415, found 1417.1446. Amino acid analysis: Ala(4) 3.8, Asp(1)1.0,Arg(1) 1.0, Gly(2) 1.9, His(1) 0.8, Pro(5) 5.2, Ser(2) 1.6, Thr(3) 2.7,Val(1)0.9.。
H-His-Gly-Val-Thr(Neu5Acα2→3Galβ1→3GalNAcα)-Ser-Ala-Pro-Asp-Thr-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(12)
化合物2と同様の条件により合成した。
凍結乾燥粉末(2 mg, 収率14 %). MALDI-TOF MS: m/z calcd for C105H169N28O45[M+H]+2542.2, found 2451.6. ESI-HRMS: m/z calcd for C105H168N28O45[M+3H]3+848.0651, found 848.0668. Amino acid analysis: Ala(4) 4.0, Asp(1)1.0,Arg(1) 1.0, Gly(2) 2.0, His(1) 0.8, Pro(5) 5.3, Ser(2) 1.7, Thr(3) 2.8,Val(1)1.0.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(13)
化合物1と同様の条件により合成した。
凍結乾燥粉末(11 mg, 収率20 %). MALDI-TOF MS: m/z calcd for C160H253N51O54[M+H]+3753.9, found 3751.1.ESI-HRMS: m/z calcd for C160H253N51O54[M+4H]4+939.2233, found 939.2245. Amino acid analysis: Ala(4) 4.1, Asp(1)1.0, Arg(1)1.0, Gly(2) 2.0, His(1) 0.9, Pro(5) 5.3, Ser(2) 1.7, Thr(3) 2.8,Val(1) 0.9.。
H-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(14)
化合物1と同様の条件により合成した。
凍結乾燥粉末(4 mg, 収率12 %). MALDI-TOF MS: m/z calcd for C264H418N79O96[M+H]+ 6231.0,found 6233.9.ESI-HRMS:m/zcalcd for C264H421N79O96[M+4H]4+1558.5123, found 1558.5117.。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Galβ1→3GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(15)
化合物1と同様の条件により合成した。
凍結乾燥粉末(5 mg, 収率 35 %). MALDI-TOF MS: m/z calcd forC114H184N31O43S[M+H]+ 2707.3, found 2707.7.ESI-HRMS:m/z calcd for C114H185N31O43SNa[M+Na+2H]3+910.4287, found 910.4279. Amino acid analysis: Ala(4) 3.9, Asp(1)1.0, Arg(1)1.0, Gly(2) 2.0, His(1) 1.2, Pro(5) 5.2, Ser(2) 1.7, Thr(3) 2.8,Val(1) 1.0. 。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(Neu5Acα2→3Galβ1→3GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(16)
化合物2と同様の条件により合成した。
凍結乾燥粉末 (3 mg, quant.). MALDI-TOF MS: m/zcalcd for C125H201N32O51S[M+H]+2998.4, found 2998.6.ESI-HRMS: m/z calcd for C125H203N32O51S[M+3H]3+1000.1332, found 1000.1322. Amino acid analysis: Ala(4)3.9, Asp(1) 1.0, Arg(1)1.0, Gly(2) 2.0, His(1) 1.2, Pro(5) 5.1, Ser(2) 1.7,Thr(3) 2.8, Val(1) 1.0.。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(17)
化合物1と同様の条件により合成した。
凍結乾燥粉末(3 mg, 収率39 %). MALDI-TOF MS: m/z calcd for C108H174N31O38S[M+H]+ 2545.2,found 2545.1.ESI-HRMS:m/zcalcd for C108H175N31O38S[M+2H]2+1273.1218, found 1273.1221. 。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(18)
化合物1と同様の条件により合成した。
凍結乾燥粉末(8 mg, 収率35 %). MALDI-TOF MS: m/z calcd for C196H312N57O70S[M+H]+ 4616.2,found 4618.3.ESI-HRMS:m/zcalcd for C196H314N57O70S[M+3H]3+1539.4161, found 1539.4167. 。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(19)
化合物1と同様の条件により合成した。
凍結乾燥粉末(8 mg, 収率24 %). MALDI-TOF MS: m/z calcd for C284H450N83O102S[M+H]+ 6687.2,found 6691.5.ESI-HRMS:m/zcalcd for C284H453N83O102S[M+4H]4+1672.5633, found 1672.5632. 。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(20)
化合物1と同様の条件により合成した。
凍結乾燥粉末(2 mg, 収率10 %). MALDI-TOF MS: m/z calcd for C372H588N109O134S[M+H]+ 8758.2,found 8763.7.ESI-HRMS:m/zcalcd for C372H591N109O134S[M+4H]4+2190.3126, found 2190.3124. 。
Biotin-PEG-linker-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-His-Gly-Val-Thr-Ser-Ala-Pro-Asp-Thr(GalNAcα)-Arg-Pro-Ala-Pro-Gly-Ser-Thr-Ala-Pro-Pro-Ala-NH2(21)
化合物1と同様の条件により合成した。
凍結乾燥粉末(2 mg, 収率7 %). MALDI-TOF MS: m/z calcd for C460H726N135O166S[M+H]+ 10830.2,found 10834.7.ESI-HRMS:m/zcalcd for C460H729N135O166S[M+4H]4+2708.0618, found 2708.0586. 。
実施例化合物の化学構造式
化合物1
(免疫原の調製)
5mgの2,3‐ST(DT*R)‐20(表1の化合物番号2)を0.2mlの蒸留水に溶解し、860μgのSulfo‐SMCC(PIERCE社製)を含む0.2mlの水溶液および0.2mlの0.1Mリン酸緩衝液(pH7.4)を加え室温にて1時間反応させた。反応液に200μgのSulfo‐SMCCを2回追加し、マレイミド化化合物2をHPLCにて精製し、凍結乾燥した。
調製した免疫原100μgをフロイント完全アジュバントと共に4週齢A/J Jms Slc雌マウスに腹腔内投与し、初回免疫とした。その後、21日後および42日後に免疫原100μgをフロイント不完全アジュバントと共に投与し、追加免疫とした。さらに71日後に免疫原100μgを生理食塩水0.1mlに縣濁した溶液を腹腔内投与し、最終免疫とした。
最終免疫の3日後に脾臓を摘出し、脾臓細胞を回収した。脾臓細胞とマウスミエローマ細胞(p3×63‐Ag8.U1、東京腫瘤研究所)を50%のポリエチレングリコール4000を用いて融合させ、ヒポキサンチン、アミノプテリン、およびチミジンを含む培地で選択した。
細胞融合10日後に特異抗体産生細胞のスクリーニングを行った。スクリーニングに用いたELSIAは以下の通りである。384穴マイクロタイタープレート(ヌンク社製)の各ウェルに0.35μgの抗マウスIgG抗体(シバヤギ社製)を含むトリス緩衝液(50mM Tris‐HCl、pH7.5)を35μl加えて4℃16時間固定した。これらのウェルを90μlの洗浄液(0.01% Tween20を含む生理食塩水)で1回洗浄した後、ブロックエース(大日本住友製薬社製)を200μl加えて室温で2時間放置して、ブロッキングを行った(抗マウスIgG抗体固相化プレート)。各ウェルを90μlの洗浄液で1回洗浄した後、10μlのハイブリドーマ培養上清と10μlの緩衝液A(0.5% ウシ血清アルブミン、0.01% Tween80、0.05% Proclin150、0.15M NaClを含む50mMトリス緩衝液、pH7.4)および0.01ngのBiotin‐2,3‐ST(DT*R)‐20(表1の化合物番号16)と2ngのStreptavidin‐HRP(PIERCE社製)を含む10μlの緩衝液Aを加え、4℃で16時間反応させた。次に各ウェルを90μlの洗浄液で3回洗浄した後に、25μlのTMB+‐Substrate‐Chromogen(DAKO社製)を添加して室温で30分間発色させた後に、25μlの0.05Mの硫酸を添加し反応を停止し、450nmにおける吸光度を測定した。スクリーニングの結果から、2,3‐ST(DT*R)‐20と強い親和性を示すクローン(1B2)を得た。マウスモノクローナル抗体アイソタイピングELISAキット(BDバイオサイエンス社製)を用いて、抗体のサブクラスを決定した結果、1B2のアイソタイプはIgG2aであった。
(糖鎖特異性)
抗マウスIgG抗体固相化プレートに、MUC1抗体を含む15μlの緩衝液Aを加え、室温で3時間反応させた。次に各ウェルを90μlの洗浄液で3回洗浄した後に、Streptavidin‐HRPとBiotin‐Tn(DT*R)‐100(表1の化合物番号21)、およびT(DT*R)‐20(化合物番号1)、2,3‐ST(DT*R)‐20(化合物番号2)、Tn(DT*R)‐20(化合物番号3)、STn(DT*R)‐20(化合物番号4)、2,3ST6G(DT*R)‐20(化合物番号6)、2,3ST6L(DT*R)‐20(化合物番号7)、2,3‐ST6SL(DT*R)‐20(化合物番号8)、C6(DT*R)‐20(化合物番号9)、ST2‐6(DT*R)‐20(化合物番号10)、dST(DT*R)‐20(化合物番号11)、2,3ST(VT*S)‐20(化合物番号12)、40(化合物番号13)をそれぞれ含む15μlの緩衝液Aを加え、4℃で16時間反応させた。次に各ウェルを90μlの洗浄液で3回洗浄した後に、15μlのTMB+−Substrate‐Chromogen(DAKO社製)を添加して室温で30分間発色させた後に、15μlの0.05Mの硫酸を添加し反応を停止し、450nmにおける吸光度を測定した。その結果、1B2は、癌細胞で高発現する糖鎖構造(T(DT*R)‐20、2,3‐ST(DT*R)‐20、Tn(DT*R)‐20)に対して高い親和性を示すが、正常細胞で高発現する糖鎖構造(2,3ST6L(DT*R)‐20、2,3‐ST6SL(DT*R)‐20)との交差反応性は低いことが示された(図1、表2)。
384穴マイクロタイタープレート(ヌンク社製)の各ウェルに0.35μgのStreptavidin(PIERCE社製)を含むトリス緩衝液(50mM Tris‐HCl、pH7.5)を35μl加えて4℃16時間固定した。これらのウェルを90μlの洗浄液(0.01% Tween20を含む生理食塩水)で1回洗浄した後、ブロックエース(大日本製薬社製)を200μl加えて室温で2時間放置して、ブロッキングを行った(Strepavidin固相化プレート)。各ウェルを90μlの洗浄液で1回洗浄した後、Biotin‐Tn(DT*R)‐20(化合物番号17)、Biotin‐Tn(DT*R)-40(化合物番号18)、Biotin‐Tn(DT*R)‐60(化合物番号19)、Biotin‐Tn(DT*R)‐80(化合物番号20)、Biotin‐Tn(DT*R)‐100(化合物番号21)をそれぞれ含む15μlの緩衝液Aを加え、室温で30分間反応させた。次に各ウェルを90μlの洗浄液で3回洗浄した後に、MUC1抗体を含む15μlの緩衝液Aを加え、4℃で16時間反応させた。次に各ウェルを90μlの洗浄液で3回洗浄した後に、15μlのTMB+−Substrate−Chromogen(DAKO社製)を添加して室温で30分間発色させた後に、15μlの0.05Mの硫酸を添加し反応を停止し、450nmにおける吸光度を測定した。その結果、1B2は、Biotin‐Tn(DT*R)‐40、Biotin‐Tn(DT*R)‐60、Biotin‐Tn(DT*R)‐80、Biotin‐Tn(DT*R)‐100との反応性はほぼ同じであり、タンデムリピートの長さ依存性が低い抗体であることが示された(図4)。一方、PankoMabは、ペプチド鎖が長い程、反応性が強いことから、タンデムリピートの長さ依存性が高い抗体であることが示された。したがって、本発明の抗体は、Tn−100マービオチンに対する親和性が、1.0×10−9(M)よりも低いものであるといえ、Tn20マータンデム構造断片に対して結合する能力を有するものであるといえる。より詳細には、本発明の抗体は、Tn20マービオチンを用いた場合の450nmの吸光度(A20)と、Tn100マービオチンを用いた場合の450nmの吸光度(A100)の比(A100/A20)が2以下であるともいえる。
センサーチップSA(GEヘルスケア社製)にBiotin‐Tn(DT*R)‐100(化合物番号21)を固相化し、以下のMUC1抗体、1B2、現在前臨床段階にあるPankoMab(GLYCOPOPE社製)、糖ペプチド免疫により得られた抗体であるVU‐2G7(MONOSAN社製)、0.N.272(SantaCruz社製)、C595(Acris社製)、B416(GeneTex社製)、VU‐3C6(Exalpha Biologicals社製)についてそれぞれの解離定数をBiacoreT100を用いて解析した結果、以下の表3の通りであった。表に示すとおり、解離定数KDは1B2が3.7×10−10(M)、17H2が2.2×10−10(M)を示し、その他の抗体と比較して小さかった。
ホルマリン固定し、パラフィン包埋したヒト乳癌組織の切片および乳癌周辺正常組織(BioCahin社)に対する1B2抗体の免疫組織染色をVectastain elite ABC kit(Vector Laboratories社製)とDAB(Roche社製)を用いて添付マニュアルに従い実施した。その結果、マウスIgG2a抗体(SIGMA−ALDRICH社製)は、乳癌組織および乳癌周辺正常組織の両方で染色像を認められなかったのに対して、1B2は、乳癌組織に染色像が認められたが、乳癌周辺正常組織では染色像はほとんど認められなかった(図5)。このことから、抗体1B2は、正常組織とは結合せず、癌組織に結合することが示された。
本実施例では、実施例2で作製した抗体について、その抗体依存性細胞傷害活性を調べた。
細胞傷害活性(cytotoxicity,%)=(実験値−自然遊離)/(最大遊離−自然遊離)×100
その結果、1B2によって約15%の細胞傷害活性が誘導された。
癌細胞膜表面に発現するMUC1タンパク質と1B2抗体が結合するかどうかを、FACSにて調べた。Trypsin(Invitrogen社製)で剥がした、ヒト乳癌細胞株T−47D(ATCC、HTB−133)およびヒト乳腺上皮細胞184A1(ATCC、CRL−8798)を、FACS buffer(5% FCS、0.05% アジ化ナトリウムを含むPBS)で1回洗浄後に、10μg/mlの1B2抗体を含む100μlのFACS bufferで縣濁し、室温で2時間反応させる。FACS bufferにて2回洗浄した後に200μlのFITC−Goat Anti−Mouse IgG(ZYMED社製)で縣濁し、室温で1時間反応させる。FACS bufferにて2回洗浄した後に、FACSAria(BD社製)で解析した。その結果、T−47Dでは、コントロールマウスIgG2a抗体(SIGMA−ALDRICH社製)と比較して、1B2抗体はFACSの蛍光シグナルが大きくシフトし(約300倍)、一方、184Aでは、1B2による蛍光シフトはほとんどなかった(約2倍)。この結果から、1B2抗体は、乳癌細胞と強く反応するが、乳腺上皮細胞とは、ほとんど反応しないことが示された。
次に、1B2について、常法を用いて可変領域の配列を決定した。その手法は以下のとおりである。
MUC1を定量するサンドイッチイムノアッセイを以下の方法で行った。
配列番号2:抗体1B2の重鎖可変領域のアミノ酸配列
配列番号3:抗体1B2の軽鎖可変領域のアミノ酸配列
配列番号4:抗体1B2のCDR1のアミノ酸配列
配列番号5:抗体1B2のCDR2のアミノ酸配列
配列番号6:抗体1B2のCDR3のアミノ酸配列
配列番号7:抗体1B2のCDR1’のアミノ酸配列
配列番号8:抗体1B2のCDR2’のアミノ酸配列
配列番号9:抗体1B2のCDR3’のアミノ酸配列
配列番号10:Panko1の重鎖可変領域のアミノ酸配列
配列番号11:Panko1の軽鎖可変領域のアミノ酸配列
配列番号12:Panko2の重鎖可変領域のアミノ酸配列
配列番号13:Panko2の軽鎖可変領域のアミノ酸配列
配列番号14:抗体1B2の重鎖可変領域の全長のアミノ酸配列
配列番号15:抗体1B2の軽鎖可変領域の全長のアミノ酸配列
Claims (8)
- 免疫グロブリン重鎖可変領域(VH)ドメインおよび免疫グロブリン軽鎖可変領域(VL)ドメインを含む少なくとも1つの抗原結合部位を有しており、該重鎖可変領域ドメインは、その配列中に、超可変領域CDR1、CDR2、CDR3を含み、CDR1は、NYGLS(配列番号4)という配列からなり、CDR2は、ENHPGSGIIYHNEKFRG(配列番号5)という配列からなり、CDR3は、SSGTRGFAY(配列番号6)という配列からなり、
該軽鎖可変領域ドメインは、その配列中に、超可変領域CDR1’、CDR2’、CDR3’を含み、CDR1’は、RSSQSIVHSNGNTYLE(配列番号7)という配列からなり、CDR2’は、KVSNRFS(配列番号8)という配列からなり、CDR3’は、FQGSHGPWT(配列番号9)という配列からなる、抗体、またはその抗原結合性断片であって、該抗体、またはその抗原結合性断片のMUC1のがん関連構造に対する特異性が、MUC1の正常組織関連構造に対するものに比べ100倍以上であり、該正常組織関連構造は、Neu5Acα2→3Galβ1→3[Galβ1→4GlcNAcβ1→6]GalNAcα−RおよびNeu5Acα2→3Galβ1→3[Neu5Acα2→3Galβ1→4GlcNAcβ1→6]GalNAcα−Rからなる群より選択され、該がん関連構造は、Neu5Acα2→3Galβ1→3GalNAcα−R、GalNAcα−RおよびGalβ1→3GalNAcα−Rからなる群より選択され、ここで、Neu5AcはN−アセチルノイラミン酸であり、Galはガラクトースであり、GlcNAcは、N−アセチルグルコサミンであり、GalNAcはN−アセチルガラクトサミンであり、Rは非糖部分である、抗体、またはその抗原結合性断片。 - 配列番号2もしくは14および3もしくは15を含むアミノ酸配列を有する、請求項1に記載の抗体、またはその抗原結合性断片。
- 請求項1または2に記載の抗体、またはその抗原結合性断片を含む医薬組成物。
- 抗がん剤である、請求項3に記載の医薬組成物。
- 請求項1または2に記載の抗体、またはその抗原結合性断片を含む、診断キット。
- 請求項1または2に記載の抗体、またはその抗原結合性断片をコードする核酸分子。
- 標識した、請求項1または2に記載の抗体、またはその抗原結合性断片。
- 請求項1〜2および7のいずれか1項に記載の抗体、またはその抗原結合性断片を用いる、イムノアッセイ。
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Families Citing this family (33)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8518405B2 (en) | 2009-10-08 | 2013-08-27 | The University Of North Carolina At Charlotte | Tumor specific antibodies and uses therefor |
EP2560989B1 (en) | 2010-04-19 | 2017-03-22 | Sumitomo Bakelite Co., Ltd. | Cancer-related glycopeptide epitopes, antibodies and methods of use |
EP2565268A4 (en) * | 2010-04-28 | 2013-10-09 | Shionogi & Co | NEW MUC1 ANTIBODY |
WO2011145085A2 (en) * | 2010-05-21 | 2011-11-24 | Procognia (Israel) Ltd | Novel antibodies and methods of use for the treatment and diagnosis of cancer |
WO2012000994A1 (en) * | 2010-06-29 | 2012-01-05 | C.N.R.S. | Llt-1 antibodies with new functional properties |
EP2407487A1 (en) * | 2010-07-14 | 2012-01-18 | F-Star Biotechnologische Forschungs - und Entwicklungsges. M.B.H. | Multispecific modular antibody |
US9845362B2 (en) | 2010-10-08 | 2017-12-19 | The University Of North Carolina At Charlotte | Compositions comprising chimeric antigen receptors, T cells comprising the same, and methods of using the same |
KR20140051249A (ko) * | 2011-06-16 | 2014-04-30 | 도오쿄 인스티튜드 오브 테크놀로지 | 3''황산화코어1당쇄에 결합하는 프로브를 사용하는 뮤신1의 분석 방법, 및 유방암의 검출 또는 모니터링 방법 |
CN102621324A (zh) * | 2012-04-05 | 2012-08-01 | 武汉康迈生物技术有限公司 | 一种结直肠癌的免疫组织化学染色检测试剂盒及应用 |
CN108175856B (zh) * | 2012-08-14 | 2024-01-26 | 米纳瓦生物技术公司 | 干细胞增强疗法 |
CN103848890B (zh) * | 2012-11-30 | 2015-09-09 | 北京市结核病胸部肿瘤研究所 | Muc1自身抗体识别的抗原多肽 |
US9587032B2 (en) * | 2013-06-12 | 2017-03-07 | The Board Of Trustees Of The Leland Stanford Junior University | IgE antibodies for the inhibition of tumor metastasis |
US10208125B2 (en) * | 2013-07-15 | 2019-02-19 | University of Pittsburgh—of the Commonwealth System of Higher Education | Anti-mucin 1 binding agents and uses thereof |
EP3127919A4 (en) * | 2014-02-06 | 2017-08-16 | Medicinal Chemistry Pharmaceuticals Co., Ltd. | Antibody against mucin 4 (muc 4) glycopeptide and use therefor |
AU2015254257B2 (en) * | 2014-04-28 | 2021-02-25 | Medicinal Chemistry Pharmaceuticals, Co., Ltd. | Anti-MUC1 antibody or antigen-binding fragment of same, and use thereof |
WO2016026143A1 (en) | 2014-08-22 | 2016-02-25 | Huiru Wang | Saccharide-based biomarkers and therapeutics |
US11554181B2 (en) | 2014-09-05 | 2023-01-17 | The University Of North Carolina At Charlotte | Tumor specific antibody conjugates and uses therefor |
CN104198707B (zh) * | 2014-09-12 | 2016-09-14 | 范飞舟 | N-乙酰氨基葡萄糖在制备检测肿瘤的试剂盒中的应用 |
CN104267185B (zh) * | 2014-09-12 | 2016-05-18 | 范飞舟 | 检测肿瘤的试剂盒及其专用识别n-乙酰氨基葡萄糖的物质 |
EP3244924B1 (en) * | 2015-01-15 | 2021-04-07 | Oncoquest Pharmaceuticals Inc. | Methods of increasing delivery of anti-cancer agents to targets |
CA2976089A1 (en) | 2015-02-10 | 2016-08-18 | Minerva Biotechnologies Corporation | Humanized anti-muc1* antibodies |
EP3334757A4 (en) * | 2015-08-14 | 2019-04-03 | Merck Sharp & Dohme Corp. | ANTI-TIGIT ANTIBODIES |
JP7089470B2 (ja) * | 2015-12-02 | 2022-06-22 | アジェナス インコーポレイテッド | 抗体およびその使用方法 |
US20200123265A1 (en) * | 2015-12-02 | 2020-04-23 | Agenus Inc. | Anti-gitr antibodies and methods of use thereof |
WO2017139975A1 (en) * | 2016-02-19 | 2017-08-24 | Huiru Wang | Antibodies against n-acetylglucosamine and n-acetyl-galactosamine |
CA3043816A1 (en) | 2016-11-18 | 2018-05-24 | Astellas Pharma Inc. | Novel anti-human muc1 antibody fab fragment |
TWI795415B (zh) | 2017-07-07 | 2023-03-11 | 日商安斯泰來製藥股份有限公司 | 新穎的抗人類CEACAM5抗體Fab片段 |
US11161911B2 (en) | 2017-10-23 | 2021-11-02 | Go Therapeutics, Inc. | Anti-glyco-MUC1 antibodies and their uses |
JP7358367B2 (ja) * | 2017-10-24 | 2023-10-10 | ジーオー セラピューティクス,インコーポレイテッド | 抗グリコmuc1抗体およびその使用 |
CA3100317A1 (en) * | 2018-05-17 | 2019-11-21 | Astellas Pharma Inc. | Complex having anti-human muc1 antibody fab fragment, peptide linker and/or ligand |
KR20210013107A (ko) | 2018-05-18 | 2021-02-03 | 다이이찌 산쿄 가부시키가이샤 | 항-muc1 항체-약물 접합체 |
JOP20210180A1 (ar) | 2019-01-07 | 2023-01-30 | Astellas Pharma Inc | مادة اقتران تشتمل على ربيطة، فاصل، رابط ببتيدي وجزيء حيوي |
CA3216563A1 (en) * | 2021-04-30 | 2022-11-03 | Beatriz Aranda-Orgilles | New anti-muc1 cars and gene edited immune cells for solid tumors cancer immunotherapy |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2002502621A (ja) * | 1998-02-13 | 2002-01-29 | バステルト・ギュンター | 乳腫瘍を伴うムチンに対する特異性のある抗体、その生産方法及び使用方法 |
US20060292643A1 (en) * | 2003-01-23 | 2006-12-28 | Steffen Goletz | Recognition molecules for the treatment and detection of tumours |
Family Cites Families (18)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4208479A (en) * | 1977-07-14 | 1980-06-17 | Syva Company | Label modified immunoassays |
GB8607679D0 (en) | 1986-03-27 | 1986-04-30 | Winter G P | Recombinant dna product |
GB9009548D0 (en) | 1990-04-27 | 1990-06-20 | Celltech Ltd | Chimeric antibody and method |
KR100272077B1 (ko) | 1990-08-29 | 2000-11-15 | 젠팜인터내셔날,인코포레이티드 | 이종 항체를 생산할 수 있는 전이유전자를 가진 인간이외의 동물 |
US5545806A (en) | 1990-08-29 | 1996-08-13 | Genpharm International, Inc. | Ransgenic non-human animals for producing heterologous antibodies |
EP0940468A1 (en) * | 1991-06-14 | 1999-09-08 | Genentech, Inc. | Humanized antibody variable domain |
AU683413B2 (en) | 1992-04-13 | 1997-11-13 | Dana-Farber Cancer Institute, Inc. | Antibodies specific for carcinoma-associated antigens |
US5648267A (en) | 1992-11-13 | 1997-07-15 | Idec Pharmaceuticals Corporation | Impaired dominant selectable marker sequence and intronic insertion strategies for enhancement of expression of gene product and expression vector systems comprising same |
US5359681A (en) | 1993-01-11 | 1994-10-25 | University Of Washington | Fiber optic sensor and methods and apparatus relating thereto |
KR100569469B1 (ko) | 1997-05-14 | 2006-04-07 | 사이트릭스 시스템스, 인크. | 서버와 클라이언트노드 사이의 접속을 관리하기 위한시스템 및 방법 |
US6355244B1 (en) * | 1997-11-17 | 2002-03-12 | University Of Kentucky Research Foundation | Methods and compositions for the treatment of psoriasis |
WO2001012217A1 (en) | 1999-08-18 | 2001-02-22 | Altarex Corp. | Therapeutic antibody against muc-1 antigen and methods for their use |
US6716966B1 (en) * | 1999-08-18 | 2004-04-06 | Altarex Corp. | Therapeutic binding agents against MUC-1 antigen and methods for their use |
CA2427858A1 (en) * | 2000-11-03 | 2002-05-10 | University Of Vermont And State Agricultural College | Compositions for inhibiting grb7 |
AU2003291135A1 (en) * | 2002-11-26 | 2004-06-18 | Expressive Constructs, Inc. | Methods, biosensors and kits for detecting and identifying fungi |
US7393531B2 (en) * | 2003-01-21 | 2008-07-01 | Arius Research Inc. | Cytotoxicity mediation of cells evidencing surface expression of MCSP |
UY28886A1 (es) * | 2004-05-10 | 2005-12-30 | Boehringer Ingelheim Int | Anticuerpos que consisten en polipéptidos y derivados conprendiendo tres secuencias conteniendo respectivamente los siguientes números de seq. id: 1-3 y 4-6; 7-9 y 10-12 y 13-15 ó 16-18 |
JP2008277344A (ja) | 2007-04-25 | 2008-11-13 | Nichicon Corp | 固体電解コンデンサの製造方法 |
-
2009
- 2009-10-28 CA CA2741798A patent/CA2741798A1/en not_active Abandoned
- 2009-10-28 WO PCT/JP2009/068531 patent/WO2010050528A1/ja active Application Filing
- 2009-10-28 JP JP2010535828A patent/JP5773352B2/ja active Active
- 2009-10-28 US US13/126,745 patent/US8722856B2/en not_active Expired - Fee Related
- 2009-10-28 CN CN200980153871.4A patent/CN102264765B/zh not_active Expired - Fee Related
- 2009-10-28 EP EP09823638.3A patent/EP2351777B1/en not_active Not-in-force
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2002502621A (ja) * | 1998-02-13 | 2002-01-29 | バステルト・ギュンター | 乳腫瘍を伴うムチンに対する特異性のある抗体、その生産方法及び使用方法 |
US20060292643A1 (en) * | 2003-01-23 | 2006-12-28 | Steffen Goletz | Recognition molecules for the treatment and detection of tumours |
Non-Patent Citations (4)
Title |
---|
JPN6014018183; Glycobiology Vol.14, No.8, 2004, p.681-692 * |
JPN6014018186; Glycobiology Vol.17, No.2, 2007, p.197-209 * |
JPN6014018187; Journal of Immunological Methods Vol.270, 2002, p.199-209 * |
JPN6014018190; Cancer Immunol Immunother Vol.55, 2006, p.1337-1347 * |
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Free format text: JAPANESE INTERMEDIATE CODE: R250 |
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R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
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R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |