JPH0549270B2 - - Google Patents
Info
- Publication number
- JPH0549270B2 JPH0549270B2 JP59235845A JP23584584A JPH0549270B2 JP H0549270 B2 JPH0549270 B2 JP H0549270B2 JP 59235845 A JP59235845 A JP 59235845A JP 23584584 A JP23584584 A JP 23584584A JP H0549270 B2 JPH0549270 B2 JP H0549270B2
- Authority
- JP
- Japan
- Prior art keywords
- urokinase
- crude
- purifying
- mmho
- conductivity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 102000003990 Urokinase-type plasminogen activator Human genes 0.000 claims description 56
- 108090000435 Urokinase-type plasminogen activator Proteins 0.000 claims description 56
- 229960005356 urokinase Drugs 0.000 claims description 56
- 238000000034 method Methods 0.000 claims description 25
- 239000001913 cellulose Substances 0.000 claims description 13
- 229920002678 cellulose Polymers 0.000 claims description 13
- -1 sulfopropyl Chemical group 0.000 claims description 11
- 210000002700 urine Anatomy 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 4
- 239000012531 culture fluid Substances 0.000 claims description 2
- 239000000243 solution Substances 0.000 description 11
- 239000008363 phosphate buffer Substances 0.000 description 7
- 230000000694 effects Effects 0.000 description 5
- 239000012535 impurity Substances 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000001179 sorption measurement Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 239000003463 adsorbent Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 239000003527 fibrinolytic agent Substances 0.000 description 2
- 230000003480 fibrinolytic effect Effects 0.000 description 2
- 239000005373 porous glass Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 239000012266 salt solution Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- DPEYHNFHDIXMNV-UHFFFAOYSA-N (9-amino-3-bicyclo[3.3.1]nonanyl)-(4-benzyl-5-methyl-1,4-diazepan-1-yl)methanone dihydrochloride Chemical compound Cl.Cl.CC1CCN(CCN1Cc1ccccc1)C(=O)C1CC2CCCC(C1)C2N DPEYHNFHDIXMNV-UHFFFAOYSA-N 0.000 description 1
- NLHHRLWOUZZQLW-UHFFFAOYSA-N Acrylonitrile Chemical compound C=CC#N NLHHRLWOUZZQLW-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 1
- 206010003178 Arterial thrombosis Diseases 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 239000005909 Kieselgur Substances 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 102000013566 Plasminogen Human genes 0.000 description 1
- 108010051456 Plasminogen Proteins 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 229920002873 Polyethylenimine Polymers 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 230000001174 ascending effect Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 239000000440 bentonite Substances 0.000 description 1
- 229910000278 bentonite Inorganic materials 0.000 description 1
- SVPXDRXYRYOSEX-UHFFFAOYSA-N bentoquatam Chemical compound O.O=[Si]=O.O=[Al]O[Al]=O SVPXDRXYRYOSEX-UHFFFAOYSA-N 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 229910052588 hydroxylapatite Inorganic materials 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011403 purification operation Methods 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000012209 synthetic fiber Substances 0.000 description 1
- 229920002994 synthetic fiber Polymers 0.000 description 1
- 239000003104 tissue culture media Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はウロキナーゼの精製法に関し、特に好
ましくは、人尿またはそれに由来する粗製ウロキ
ナーゼ、組織培養液またはそれに由来する粗製ウ
ロキナーゼ等の大量の水溶液中から短時間で効率
的に精製する方法に関する。Detailed Description of the Invention [Industrial Application Field] The present invention relates to a method for purifying urokinase, and particularly preferably a large amount of aqueous solution such as human urine or crude urokinase derived therefrom, tissue culture medium or crude urokinase derived therefrom. This invention relates to a method for efficiently purifying medium to medium-sized particles in a short period of time.
〔従来技術〕
ウロキナーゼは血清中に含まれるプラスミノー
ゲンを活性化して、フイブリン溶解能を有するプ
ラスミンを生成する機能があるため、線溶系の賦
活剤として有用であり、末梢動脈血栓症や心筋梗
塞症などの治療に広く臨床使用されている。又、
ウロキナーゼは近年抗癌剤としての用途も開発さ
れ、その需要は急増している。[Prior art] Urokinase has the function of activating plasminogen contained in serum to generate plasmin with fibrinolytic ability, so it is useful as an activator of the fibrinolytic system, and is useful for peripheral arterial thrombosis and myocardial infarction. It is widely used clinically to treat diseases such as or,
Urokinase has recently been developed for use as an anticancer agent, and its demand is rapidly increasing.
ウロキナーゼの精製法としては、燐酸セルロー
ス、アンバーライトIRC−50、カルボキシメチル
デキストラン、カルボキシメチルセルロース、ス
ルホエチルセルロース等のイオン交換体を用いる
方法、シリカゲル、ガラスビーズ、多孔性ガラ
ス、カーボワツクス多孔性ガラス、珪藻土、活性
化珪藻土、ハイドロキシアパタイト、ベントナイ
ト等の物理化学的吸着剤を用いる方法、アクリル
ニトリル合成繊維、シアノアルキル化セルロー
ス、ポリエチレンイミン含有ポリアミド繊維、寒
天、アガロース等の特異的吸着剤を用いる方法な
ど、種々の方法が知られている。 Methods for purifying urokinase include methods using ion exchangers such as cellulose phosphate, Amberlite IRC-50, carboxymethyl dextran, carboxymethyl cellulose, and sulfoethyl cellulose, silica gel, glass beads, porous glass, carbo wax porous glass, diatomaceous earth, Various methods include methods using physicochemical adsorbents such as activated diatomaceous earth, hydroxyapatite, and bentonite, and methods using specific adsorbents such as acrylonitrile synthetic fibers, cyanoalkylated cellulose, polyamide fibers containing polyethyleneimine, agar, and agarose. method is known.
本発明の目的は、短時間で効率がよくウロキナ
ーゼを精製する方法を提供することである。
An object of the present invention is to provide a method for purifying urokinase in a short time and with high efficiency.
本発明者らは、効率の良いウロキナーゼの精製
法について種々研究を重ねた結果、特定の条件下
においてスルホプロピルセルロースがウロキナー
ゼに対して特異的な吸着作用を有していることを
見出し、かかる知見に基づいて、粗ウロキナーゼ
溶液から発熱性物質、不純物（例えば、色素、に
ごり、不純蛋白等）等を除去し、ウロキナーゼを
効率良く、しかも経済的に収得する方法を確立す
るに至つた。
As a result of various studies on efficient purification methods for urokinase, the present inventors discovered that sulfopropyl cellulose has a specific adsorption effect on urokinase under certain conditions, and based on this knowledge, Based on this, we have established a method for efficiently and economically obtaining urokinase by removing pyrogens, impurities (e.g., pigments, cloudiness, impure proteins, etc.) from crude urokinase solutions.
即ち、本発明はウロキナーゼをスルホプロピル
セルロースに吸着させることからなる粗製ウロキ
ナーゼからのウロキナーゼの精製方法において、
ウロキナーゼをスルホプロピルセルロースに吸着
させる前に粗製ウロキナーゼ溶液およびスルホプ
ロピルセルロースを電導度30ｍmho／cm以下とな
るように調整しておき、かつ電導度40ｍmho／cm
以上の条件下で吸着したウロキナーゼを溶出させ
ることを特徴とするウロキナーゼの精製方法であ
る。 That is, the present invention provides a method for purifying urokinase from crude urokinase, which comprises adsorbing urokinase to sulfopropylcellulose,
Before adsorbing urokinase to sulfopropyl cellulose, the crude urokinase solution and sulfopropyl cellulose are adjusted to have an electrical conductivity of 30 mmho/cm or less, and the electrical conductivity is 40 mmho/cm.
This is a method for purifying urokinase, which is characterized by eluting adsorbed urokinase under the above conditions.
以下、本発明方法を具体的に説明する。 The method of the present invention will be specifically explained below.
本発明における出発原料である粗製ウロキナー
ゼは、水溶液状態として本発明の吸着操作に付さ
れる。粗製ウロキナーゼとしては、好ましくは人
尿またはそれに由来する粗製ウロキナーゼ、組織
培養液またはそれに由来する粗製ウロキナーゼが
あげられる。 Crude urokinase, which is the starting material in the present invention, is subjected to the adsorption operation of the present invention in the form of an aqueous solution. The crude urokinase preferably includes human urine or crude urokinase derived therefrom, tissue culture fluid or crude urokinase derived therefrom.
本発明で使用されるスルホプロピルセルロース
は、好ましくはシート状素材を渦巻状に巻いた態
様で本発明の精製操作に付される。かかる渦巻状
で巻いた態様（以下、カートリツジタイプとい
う）のものとしては、例えばAMF社のZeta
prep
等のカートリツジタイプのものがあげら
れる。第１図はカートリツジタイプの一実施例の
垂直断面図であり、入口１から入つた粗製ウロキ
ナーゼ溶液は、外周から内側へ向かい中央のコア
２に集められ、出口３より出る。矢印は粗製ウロ
キナーゼ溶液の流れを示すものである。４はベン
トを、５はドレインを示す。第２図は横断面図で
あり、外周から入つた粗製ウロキナーゼ溶液がコ
アに集められている状態を示すものである。 The sulfopropylcellulose used in the present invention is preferably subjected to the purification operation of the present invention in the form of a spirally wound sheet material. Examples of such a spirally wound type (hereinafter referred to as a cartridge type) include AMF's Zeta.
Cartridge type products such as prep are listed. FIG. 1 is a vertical cross-sectional view of an embodiment of the cartridge type, in which the crude urokinase solution enters from the inlet 1, is collected in the central core 2 from the outer periphery inward, and exits from the outlet 3. Arrows indicate the flow of crude urokinase solution. 4 indicates a vent, and 5 indicates a drain. FIG. 2 is a cross-sectional view showing a state in which the crude urokinase solution entered from the outer periphery is collected in the core.
本発明においては、電導度を30ｍmho／cm以下
に調整した粗製ウロキナーゼ溶液を、同じく予め
電導度を30ｍmho／cm以下に調整ししておいたス
ルホプロピルセルロースに接触させることによつ
て、ウロキナーゼを特異的に吸着させる。その際
の吸着操作はPH５〜７で行うのが好ましい。この
条件でウロキナーゼはスルホプロピルセルロース
に特異的に吸着され、大部分の不純物は吸着され
ず、そのまま通過する。次いで、ウロキナーゼ吸
着スルホプロピルセルロースを洗浄液〔好ましく
は、PH３〜８の希薄塩溶液（例えば、電導度30ｍ
mho／cm以下の燐酸緩衝液）〕で洗浄する。この
洗浄操作により、更に不純物が除かれ、ウロキナ
ーゼは吸着されたまま残る。吸着しているウロキ
ナーゼは電導度40ｍmho／cm以上に調整した溶出
液によつて溶出される。溶出液としては、PH6.5
〜11の濃厚塩溶液（例えば電導度40ｍmho／cm以
上の燐酸緩衝液、食塩溶液）、アンモニア水等が
例示される。 In the present invention, urokinase can be isolated by contacting a crude urokinase solution whose conductivity has been adjusted to 30 mmho/cm or less with sulfopropyl cellulose whose conductivity has also been adjusted to 30 mmho/cm or less. to be adsorbed. At this time, the adsorption operation is preferably performed at a pH of 5 to 7. Under these conditions, urokinase is specifically adsorbed to sulfopropyl cellulose, and most impurities are not adsorbed and pass through as is. Next, the urokinase-adsorbed sulfopropyl cellulose is washed with a washing solution [preferably a dilute salt solution with a pH of 3 to 8 (for example, conductivity of 30 m
Wash with phosphate buffer (no more than mho/cm). This washing operation further removes impurities and leaves the urokinase adsorbed. The adsorbed urokinase is eluted with an eluent whose conductivity is adjusted to 40 mmho/cm or higher. As an eluent, PH6.5
-11 concentrated salt solutions (for example, phosphate buffers and saline solutions with an electrical conductivity of 40 mmho/cm or more), aqueous ammonia, and the like.
かくして得られるウロキナーゼ溶出液は、更に
精製することが好ましく、たとえば常法により硫
安塩析した後、透析することによつて、より高度
に精製されたウロキナーゼを得ることができる。 The urokinase eluate thus obtained is preferably further purified. For example, more highly purified urokinase can be obtained by salting out ammonium sulfate using a conventional method and then dialysis.
スルホプロピルセルロースは、電動度30ｍ
mho／cm以下の条件において、選択的にウロキナ
ーゼを吸着するものであり、本発明の精製方法に
よれば、大量の粗製ウロナーゼを水溶液中からウ
ロキナーゼを短時間で効率的に精製することがで
きる。特に、スルホプロピルセルロースとしてシ
ート状素材を渦巻状に巻いた態様のものを使用し
た場合、カラム法に於けるカラムへの樹脂等の充
填やバツチ法における濾過、または遠心等による
樹脂等の回収は全く不要である利点を有してお
り、非常に効率的な方法である。従つて、工業的
規膜におけるウロキナーゼの製法として極めて有
利である。
Sulfopropyl cellulose has a conductivity of 30 m.
Urokinase is selectively adsorbed under conditions of mho/cm or less, and according to the purification method of the present invention, urokinase can be efficiently purified from a large amount of crude urokinase in an aqueous solution in a short time. In particular, when using sulfopropyl cellulose in the form of a spirally wound sheet material, it is difficult to fill the column with resin in the column method, filtrate in the batch method, or collect the resin by centrifugation, etc. It has the advantage that it is not necessary at all, making it a very efficient method. Therefore, this method is extremely advantageous as a method for producing urokinase in industrial membranes.
以下、実施例を挙げて、本発明方法を説明す
る。 The method of the present invention will be explained below with reference to Examples.
なお、実施例中ウロキナーゼの力価はプロウグ
らのフイブリン平板法〔J.Plougら：Biochim.
Biophys.Acta24、278（1957）〕にて測定した。 In addition, the titer of urokinase in the examples was determined by the fibrin plate method of Ploug et al. [J. Ploug et al.: Biochim.
Biophys. Acta 24 , 278 (1957)].
実施例 １
ゼータプレツプSPカートリツジ
を水で洗浄
した後、PHを6.0に、電導度を15ｍmho／cmに調
整した尿5000を上昇法にて通過せしめる。カー
トリツジに0.01M燐酸緩衝液（PH6.5）50を流
して、カートリツジに吸着した不純物を流出除去
せしめる。Example 1 After washing a Zetaprep SP cartridge with water, 5,000 ml of urine whose pH was adjusted to 6.0 and conductivity to 15 mmho/cm was passed through in an ascending manner. 50 g of 0.01M phosphate buffer (PH6.5) is poured into the cartridge to remove impurities adsorbed to the cartridge.
次いで、カートリツジに、0.8M食塩を含んだ
0.01M燐酸緩衝液（PH8.0）５を流してウロキ
ナーゼを溶出させる。このようにして得られたウ
ロキナーゼ溶液の比活性は8000単位／mg蛋白であ
り、また精製により85000単位／mg蛋白のウロキ
ナーゼを得ることが出来た。また、尿からのウロ
キナーゼの回収率は90％あつた。 Next, the cartridge contained 0.8M salt.
Flow 0.01M phosphate buffer (PH8.0) 5 to elute urokinase. The specific activity of the urokinase solution thus obtained was 8,000 units/mg protein, and 85,000 units/mg protein of urokinase could be obtained by purification. In addition, the recovery rate of urokinase from urine was 90%.
実施例 ２
0.1M燐酸緩衝液（PH6.5）で十分平衡化したゼ
ータプレツプSPカートリツジ
に比活性6500単
位／mg蛋白の粗ウロキナーゼ5000万単位を通過せ
しめる。次いでカートリツジに0.1M燐酸緩衝液
（PH6.5）を流下し、カートリツジに吸着した不純
物を流出除去せしめる。Example 2 50 million units of crude urokinase with a specific activity of 6500 units/mg protein are passed through a Zetaprep SP cartridge that has been sufficiently equilibrated with 0.1M phosphate buffer (PH6.5). Next, 0.1M phosphate buffer (PH6.5) is poured into the cartridge to remove impurities adsorbed to the cartridge.
次いで、カートリツジに、1.0M食塩を含んだ
0.1M燐酸緩衝液（PH8.0）を流下してウロキナー
ゼを溶出させる。このようにして得られたウロキ
ナーゼ溶液の比活性は55000単位／mg蛋白であり、
そのときのウロキナーゼの回収率は87％であつ
た。 Next, the cartridge contained 1.0M salt.
Urokinase is eluted by flowing down 0.1M phosphate buffer (PH8.0). The specific activity of the urokinase solution thus obtained was 55,000 units/mg protein;
The recovery rate of urokinase at that time was 87%.
第１図は、シート状素材を渦巻状に巻いた態様
のスルホプロピルセルロースの一実施例の垂直断
面図であり、第２図は、その横断面図である。
１……入口、２……中央のコア、３……出口、
４……ベント、５……ドレイン、Ａ……粗製ウロ
キナーゼ溶液の流れ。
FIG. 1 is a vertical cross-sectional view of an embodiment of sulfopropyl cellulose in the form of a spirally wound sheet material, and FIG. 2 is a cross-sectional view thereof. 1...Entrance, 2...Central core, 3...Exit,
4...Vent, 5...Drain, A...Flow of crude urokinase solution.
Claims (1)
吸着させることからなる粗製ウロキナーゼからの
ウロキナーゼの精製方法において、ウロキナーゼ
をスルホプロピルセルロースに吸着させる前に粗
製ウロキナーゼ溶液およびスルホプロピルセルロ
ースを電導度30ｍmho／cm以下となるように調整
しておき、かつ電導度40ｍmho／cm以上の条件下
で吸着したウロキナーゼを溶出させることを特徴
とするウロキナーゼの精製方法。 ２ スルホプロピルセルロースがシート状素材渦
巻状に巻いた構造物の態様である特許請求の範囲
第１項に記載のウロキナーゼの精製方法。 ３ ウロキナーゼが人尿由来または組織培養液由
来のものであることを特徴とする特許請求の範囲
第１項または第２項に記載のウロキナーゼの精製
方法。[Claims] 1. A method for purifying urokinase from crude urokinase, which comprises adsorbing urokinase onto sulfopropylcellulose, in which the crude urokinase solution and sulfopropylcellulose are heated to a conductivity of 30 mmho/min before adsorbing urokinase onto sulfopropylcellulose. 1. A method for purifying urokinase, which comprises eluating adsorbed urokinase under conditions where the conductivity is adjusted to 40 mmho/cm or less and the conductivity is 40 mmho/cm or more. 2. The method for purifying urokinase according to claim 1, wherein the sulfopropyl cellulose is a spirally wound sheet-like material. 3. The method for purifying urokinase according to claim 1 or 2, wherein the urokinase is derived from human urine or tissue culture fluid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23584584A JPS61115492A (en) | 1984-11-07 | 1984-11-07 | Method of purifying urokinase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23584584A JPS61115492A (en) | 1984-11-07 | 1984-11-07 | Method of purifying urokinase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS61115492A JPS61115492A (en) | 1986-06-03 |
| JPH0549270B2 true JPH0549270B2 (en) | 1993-07-23 |
Family
ID=16992114
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP23584584A Granted JPS61115492A (en) | 1984-11-07 | 1984-11-07 | Method of purifying urokinase |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS61115492A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS51148088A (en) * | 1975-06-16 | 1976-12-18 | Mitsubishi Rayon Co Ltd | Process for obtaining substances produced by organisms |
| GB1556652A (en) * | 1977-06-03 | 1979-11-28 | Sumitomo Chemical Co | Process for purifying urokinase |
| JPS5713270A (en) * | 1980-06-27 | 1982-01-23 | Kita Nippon Riyuutai Kiki Hanbai Kk | Combined controller and alarm for starting of automotive engine |
| JPS5935A (en) * | 1982-06-24 | 1984-01-05 | オリンパス光学工業株式会社 | Glare preventing apparatus of endoscope using liquid crystal light blocking plate |
-
1984
- 1984-11-07 JP JP23584584A patent/JPS61115492A/en active Granted
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS51148088A (en) * | 1975-06-16 | 1976-12-18 | Mitsubishi Rayon Co Ltd | Process for obtaining substances produced by organisms |
| GB1556652A (en) * | 1977-06-03 | 1979-11-28 | Sumitomo Chemical Co | Process for purifying urokinase |
| JPS5713270A (en) * | 1980-06-27 | 1982-01-23 | Kita Nippon Riyuutai Kiki Hanbai Kk | Combined controller and alarm for starting of automotive engine |
| JPS5935A (en) * | 1982-06-24 | 1984-01-05 | オリンパス光学工業株式会社 | Glare preventing apparatus of endoscope using liquid crystal light blocking plate |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS61115492A (en) | 1986-06-03 |
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