JPH01115901A - Production of gel or hardened gel - Google Patents
Production of gel or hardened gelInfo
- Publication number
- JPH01115901A JPH01115901A JP27169287A JP27169287A JPH01115901A JP H01115901 A JPH01115901 A JP H01115901A JP 27169287 A JP27169287 A JP 27169287A JP 27169287 A JP27169287 A JP 27169287A JP H01115901 A JPH01115901 A JP H01115901A
- Authority
- JP
- Japan
- Prior art keywords
- gel
- sodium alginate
- solution
- barium
- ions
- 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.)
- Granted
Links
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000000661 sodium alginate Substances 0.000 claims abstract description 21
- 235000010413 sodium alginate Nutrition 0.000 claims abstract description 21
- 229940005550 sodium alginate Drugs 0.000 claims abstract description 21
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 claims abstract description 18
- 229910001424 calcium ion Inorganic materials 0.000 claims abstract description 18
- 229910001422 barium ion Inorganic materials 0.000 claims abstract description 15
- 102000004190 Enzymes Human genes 0.000 claims abstract description 10
- 108090000790 Enzymes Proteins 0.000 claims abstract description 10
- 210000004102 animal cell Anatomy 0.000 claims abstract description 8
- 210000004027 cell Anatomy 0.000 claims abstract description 6
- 244000005700 microbiome Species 0.000 claims abstract description 6
- 239000013543 active substance Substances 0.000 claims description 8
- 238000005728 strengthening Methods 0.000 claims description 4
- 238000000465 moulding Methods 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 abstract description 14
- 229910000147 aluminium phosphate Inorganic materials 0.000 abstract description 7
- AEMOLEFTQBMNLQ-UHFFFAOYSA-N beta-D-galactopyranuronic acid Natural products OC1OC(C(O)=O)C(O)C(O)C1O AEMOLEFTQBMNLQ-UHFFFAOYSA-N 0.000 abstract description 3
- 238000005842 biochemical reaction Methods 0.000 abstract description 3
- AEMOLEFTQBMNLQ-YBSDWZGDSA-N d-mannuronic acid Chemical compound O[C@@H]1O[C@@H](C(O)=O)[C@H](O)[C@@H](O)[C@H]1O AEMOLEFTQBMNLQ-YBSDWZGDSA-N 0.000 abstract description 3
- 239000011575 calcium Substances 0.000 abstract description 2
- 239000002253 acid Substances 0.000 abstract 1
- 239000012531 culture fluid Substances 0.000 abstract 1
- 239000000499 gel Substances 0.000 description 35
- 239000000243 solution Substances 0.000 description 18
- 235000010443 alginic acid Nutrition 0.000 description 9
- 229920000615 alginic acid Polymers 0.000 description 9
- 229910052788 barium Inorganic materials 0.000 description 6
- DSAJWYNOEDNPEQ-UHFFFAOYSA-N barium atom Chemical group [Ba] DSAJWYNOEDNPEQ-UHFFFAOYSA-N 0.000 description 6
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 5
- 229940072056 alginate Drugs 0.000 description 5
- 229960001126 alginic acid Drugs 0.000 description 4
- 239000000783 alginic acid Substances 0.000 description 4
- 150000004781 alginic acids Chemical class 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 3
- WDIHJSXYQDMJHN-UHFFFAOYSA-L barium chloride Chemical compound [Cl-].[Cl-].[Ba+2] WDIHJSXYQDMJHN-UHFFFAOYSA-L 0.000 description 3
- 229910001626 barium chloride Inorganic materials 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 108090000769 Isomerases Proteins 0.000 description 2
- 102000004195 Isomerases Human genes 0.000 description 2
- IAJILQKETJEXLJ-SQOUGZDYSA-N L-guluronic acid Chemical compound O=C[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O IAJILQKETJEXLJ-SQOUGZDYSA-N 0.000 description 2
- IWOUKMZUPDVPGQ-UHFFFAOYSA-N barium nitrate Chemical compound [Ba+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O IWOUKMZUPDVPGQ-UHFFFAOYSA-N 0.000 description 2
- ZCCIPPOKBCJFDN-UHFFFAOYSA-N calcium nitrate Chemical compound [Ca+2].[O-][N+]([O-])=O.[O-][N+]([O-])=O ZCCIPPOKBCJFDN-UHFFFAOYSA-N 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 102100032534 Adenosine kinase Human genes 0.000 description 1
- 108020000543 Adenylate kinase Proteins 0.000 description 1
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 1
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 1
- 108700016171 Aspartate ammonia-lyases Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 102100026189 Beta-galactosidase Human genes 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102100035882 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 108010003989 D-amino-acid oxidase Proteins 0.000 description 1
- 102000004674 D-amino-acid oxidase Human genes 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 108010036781 Fumarate Hydratase Proteins 0.000 description 1
- 102100036160 Fumarate hydratase, mitochondrial Human genes 0.000 description 1
- 108010036164 Glutathione synthase Proteins 0.000 description 1
- 102100034294 Glutathione synthetase Human genes 0.000 description 1
- 108010043428 Glycine hydroxymethyltransferase Proteins 0.000 description 1
- 102000002667 Glycine hydroxymethyltransferase Human genes 0.000 description 1
- 102000005548 Hexokinase Human genes 0.000 description 1
- 108700040460 Hexokinases Proteins 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 108010087702 Penicillinase Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 102000004357 Transferases Human genes 0.000 description 1
- 108090000992 Transferases Proteins 0.000 description 1
- 102100033055 Transketolase Human genes 0.000 description 1
- 108010043652 Transketolase Proteins 0.000 description 1
- 108700040099 Xylose isomerases Proteins 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- ITHZDDVSAWDQPZ-UHFFFAOYSA-L barium acetate Chemical compound [Ba+2].CC([O-])=O.CC([O-])=O ITHZDDVSAWDQPZ-UHFFFAOYSA-L 0.000 description 1
- 108010005774 beta-Galactosidase Proteins 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 229960005069 calcium Drugs 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 1
- 239000001639 calcium acetate Substances 0.000 description 1
- 235000011092 calcium acetate Nutrition 0.000 description 1
- 229960005147 calcium acetate Drugs 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- MKJXYGKVIBWPFZ-UHFFFAOYSA-L calcium lactate Chemical compound [Ca+2].CC(O)C([O-])=O.CC(O)C([O-])=O MKJXYGKVIBWPFZ-UHFFFAOYSA-L 0.000 description 1
- 239000001527 calcium lactate Substances 0.000 description 1
- 229960002401 calcium lactate Drugs 0.000 description 1
- 235000011086 calcium lactate Nutrition 0.000 description 1
- FYPVXEILSNEKOO-UHFFFAOYSA-L calcium;butanoate Chemical compound [Ca+2].CCCC([O-])=O.CCCC([O-])=O FYPVXEILSNEKOO-UHFFFAOYSA-L 0.000 description 1
- 238000005266 casting Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 229950009506 penicillinase Drugs 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- -1 tyrosidase Proteins 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、アルギン酸のゲル又は固定化ゲルを強化する
方法に関するものである。DETAILED DESCRIPTION OF THE INVENTION (Industrial Field of Application) The present invention relates to a method for strengthening alginic acid gels or immobilized gels.
更に詳細には１本発明は、カルシウムイオンで形成され
たアルギン酸のゲル又は固定化ゲルをバリウムイオンで
処理し、ゲル又は固定化ゲルを強化する方法に関するも
のである。More specifically, the present invention relates to a method of treating an alginic acid gel or immobilized gel formed with calcium ions with barium ions to strengthen the gel or immobilized gel.
本発明において強化されたゲル又は固定化ゲルは燐酸等
を含む培養液によって脱弱化させることなく、長期間の
生化学的反応等に耐え得るもので、生化学界に益すると
ころ大なるものがある。The gel or immobilized gel strengthened in the present invention can withstand long-term biochemical reactions without being weakened by culture medium containing phosphoric acid, etc., and is of great benefit to the biochemical community. .
（従来の技術）
一般に、酵素、微生物、動物細胞等をアルギン酸ナトリ
ウム溶液に添加し、カルシウムイオンを含む溶液中に、
滴下したり、紡糸したりして、ゲル化して、生化学的反
応等に利用することはよく知られている。(Prior art) Generally, enzymes, microorganisms, animal cells, etc. are added to a sodium alginate solution, and the solution contains calcium ions.
It is well known that it can be applied dropwise or spun to form a gel and used for biochemical reactions and the like.
（発明が解決しようとする問題点）
しかし、従来のアルギン酸ナトリウムとカルシラムイオ
ンを反応させて得られたゲルが反応液中に含まれる種々
の塩類の影響、或いはｐＨの変化によってゲル強度が著
しく低下したり、リン酸イオン等のキレート剤の存在に
よりゲルが溶解してしまうという問題点があった。(Problems to be Solved by the Invention) However, the gel strength of the conventional gel obtained by reacting sodium alginate and calcilam ion is significantly reduced due to the influence of various salts contained in the reaction solution or changes in pH. There have been problems in that the gel is dissolved due to the presence of chelating agents such as phosphate ions.
そこでゲルの強度を向上させるため反応液中にカルシウ
ムイオンを多量に添加することも行われるが、多量のカ
ルシウムイオンによって、酵素、微生物、動物細胞等の
生化学的活性が阻害されるという問題が生じるのである
。Therefore, in order to improve the strength of the gel, large amounts of calcium ions are added to the reaction solution, but there is a problem that large amounts of calcium ions inhibit the biochemical activities of enzymes, microorganisms, animal cells, etc. It happens.
（問題点を解決するための手段）
本発明者らは、アルギン酸ゲルを長期間の培養等に耐え
る強固なものにするために鋭意研究すした結果、カルシ
ウムイオンを用いてゲル化したアルギン酸ゲルをバリウ
ムイオンで処理することによって、強固なアルギン酸ゲ
ルを得ることに成功したものである。(Means for Solving the Problems) The present inventors have conducted extensive research to make alginate gel strong enough to withstand long-term culture, etc. As a result, the present inventors have developed an alginate gel gelled using calcium ions. By treating with barium ions, we succeeded in obtaining a strong alginate gel.
本発明は、アルギン酸ナトリウム溶液をカルシウムイオ
ンで適宜形状に成型し、次いでバリウムイオンで処理し
、強化することを特徴とする強化されたゲルの製造法で
ある。The present invention is a method for producing a reinforced gel, which is characterized in that a sodium alginate solution is shaped into an appropriate shape with calcium ions, and then treated with barium ions to strengthen it.
また、本発明は、生化学的活性体を添加したアルギン酸
ナトリウム溶液をカルシウムイオンで適宜形状に成型し
、次いでバリウムイオンで処理し、強化することを特徴
とする強化された固定化ゲルの製造法である。The present invention also provides a method for producing a strengthened immobilized gel, which comprises molding a sodium alginate solution to which a biochemically active substance has been added into an appropriate shape with calcium ions, and then treating and strengthening with barium ions. It is.
本発明に使用するアルギン酸ナトリウムはカッ藻類中に
含まれるアルギン酸を化学操作により抽出しナトリウム
塩にしたもので、市販のアルギン酸ナトリウムなどいず
れも使用できる。しかしながら、アルギン酸はＤ−マン
ヌロン酸（Ｍ）とＬ−グルロン酸（Ｇ）とから構成され
ていて、両者の成分比（Ｍ／Ｇ比）が物性に大きな影響
を与えている。The sodium alginate used in the present invention is a sodium salt obtained by extracting alginic acid contained in algae by chemical manipulation, and any commercially available sodium alginate can be used. However, alginic acid is composed of D-mannuronic acid (M) and L-guluronic acid (G), and the ratio of these two components (M/G ratio) has a great influence on the physical properties.
そして、Ｍのみからなる部分、Ｇのみからべろ部分、Ｍ
とＧが混在する部分などがあるがゲル化剤の金属イオン
と結合して強固なゲルを形成するのは主としてＧブロッ
クの働きによることが明らかとなっている。Then, a part consisting only of M, a part consisting only of G, a part consisting only of M,
Although there are parts where G and G are mixed, it is clear that it is mainly the function of the G block that forms a strong gel by combining with the metal ions of the gelling agent.
本発明では、アルギン酸を構成するＤ−マンヌロン酸（
Ｍ）とＬ−グルロン酸（Ｇ）の比、すなわちＭ／Ｇ比が
０．１〜０．８、より好ましくは０．１〜０．２のもの
を選んで用いるのがよい。In the present invention, D-mannuronic acid (
It is preferable to select and use one having a ratio of M) to L-guluronic acid (G), that is, an M/G ratio of 0.1 to 0.8, more preferably 0.1 to 0.2.
この様なＭ／Ｇ比の低いアルギン酸ナトリウムは、Ｇ含
量の高い海藻類、若しくは多く含まれている部位［茎（
幹）部］、及びとれた季節を選択して抽出調製するか、
又はＧ含量の高いものをブレンドするなどの方法によっ
て、特別に調製して得ることができる。Sodium alginate with such a low M/G ratio can be used in seaweeds with high G content or in parts that contain a large amount [stem].
Extract and prepare by selecting the season in which it was harvested, or
Alternatively, it can be specially prepared and obtained by a method such as blending one with a high G content.
Ｍ／Ｇ比が上記範囲にあるものはゲル強度が大である。When the M/G ratio is within the above range, the gel strength is high.
本発明においては、カルシウムイオンで処理したアルギ
ン酸ゲルをバリウムイオンで処理することによって、ゲ
ルを強化することを特色とするものである。The present invention is characterized in that the alginate gel treated with calcium ions is treated with barium ions to strengthen the gel.
カルシウムイオンとしては硝酸カルシウム、酢酸カルシ
ウム、乳酸カルシウム、酪酸カルシウム。Calcium ions include calcium nitrate, calcium acetate, calcium lactate, and calcium butyrate.
塩化カルシウム等が用いられる。又、バリウムイオンと
しては塩化バリウム、硝酸バリウム、酢酸バリウム等の
水溶性の塩が用いられ、特に塩化バリウムが容易に入手
できて使いやすい。Calcium chloride etc. are used. Further, as the barium ion, water-soluble salts such as barium chloride, barium nitrate, barium acetate, etc. are used, and especially barium chloride is easily available and easy to use.
具体的には、生化学的活性体を添加もしくは添加しない
アルギン酸ソーダの溶液をカルシウムイオンを含む溶液
で、粒状、繊維状等に成型した後、その成型ゲルをバリ
ウムイオンを含む溶液で処理して、燐酸等に対して強化
されるものである。Specifically, a solution of sodium alginate with or without biochemically active substances is molded into granules, fibers, etc. with a solution containing calcium ions, and then the molded gel is treated with a solution containing barium ions. , phosphoric acid, etc.
ナトリウムと置換したカルシウムの一部又は大部分がバ
リウムと置き換ったり、新たにバリウムが残ったナトリ
ウムと置換したり、更にはこれらの組合せも起り、全体
としてゲルが燐酸等に対して強化されるものと考えられ
るが。Part or most of the calcium that replaced sodium is replaced with barium, barium is newly replaced with the remaining sodium, and a combination of these also occurs, and the gel as a whole is strengthened against phosphoric acid, etc. However, it is thought that
カルシウムイオンでゲル化して、更にバリウムイオンで
処理することによって、ゲルが燐酸等に対して強化され
る理由の詳細は不明である。The details of why gelling with calcium ions and further treatment with barium ions strengthens the gel against phosphoric acid and the like are unknown.
アルギン酸ナトリウムの濃度は、０．５ （Ｗ／Ｖ）％
〜８（Ｗ／Ｖ）％濃度が良く、より好ましくは０．８（
Ｗ／Ｖ）％〜３（Ｗ／Ｖ）％濃度である。The concentration of sodium alginate is 0.5 (W/V)%
A concentration of ~8 (W/V)% is good, more preferably 0.8 (
The concentration is from 3(W/V)% to 3(W/V)%.
また、アルギン酸ナトリウム溶液に添加される生化学的
活性体としては、酵素、微生物、動物細胞、植物１胞な
どがある。Biochemically active substances added to the sodium alginate solution include enzymes, microorganisms, animal cells, plant cells, and the like.
酵素としては、アルコールデビドロゲナーゼ、Ｄ−アミ
ノ酸オキシダーゼ、カタラーゼ等の酸化還元酵素、トラ
ンスケトラーゼ、アデニレートキナーゼ、ヘキソキナー
ゼ等の転移酵素、β−ガラクトシダーゼ、ペニシリナー
ゼ、リパーゼ、エステラーゼなどの加水分解酵素、フマ
ラーゼ、アスパルターゼ、スレオニンアルドラーゼ、β
−チロシダーゼなどのリアーゼ酵素、グルコースイソメ
ラーゼ、アラニンイソメラーゼなどの異性化酵素、グル
タチオンシンターゼ、グルタミンシンターゼなどのりガ
ーゼ酵素などがあげられる。Examples of enzymes include oxidoreductases such as alcohol dehydrogenase, D-amino acid oxidase, and catalase, transferases such as transketolase, adenylate kinase, and hexokinase, and hydrolyzing enzymes such as β-galactosidase, penicillinase, lipase, and esterase. Enzymes, fumarase, aspartase, threonine aldolase, β
-Lyase enzymes such as tyrosidase, isomerase enzymes such as glucose isomerase and alanine isomerase, and glue gauze enzymes such as glutathione synthase and glutamine synthase.
又、微生物としては細菌、酵母、カビ、放線菌などの酵
素活性を有する微生物であれば特に限定されることはな
く、また、動物細胞としては、各種生理活性物質を生産
する細胞株、抗体を生産するハイブリドーマなどがあり
、更に植物細胞としは各種生理活性物質を生産する細胞
がある。Microorganisms are not particularly limited as long as they have enzymatic activity such as bacteria, yeast, molds, and actinomycetes. Animal cells include cell lines that produce various physiologically active substances and antibodies. There are hybridomas that produce bioactive substances, and plant cells that produce various physiologically active substances.
生化学的活性体は０．０１〜５０％、好ましくは０．１
〜２０％程度アルギン酸ナトリウム溶液に添加され。The biochemically active form is 0.01-50%, preferably 0.1
~20% is added to the sodium alginate solution.
混合される。ただし、動物細胞の場合は、アルギン酸ナ
トリウム溶液１＋ｍｌに対しＩ Ｘ １０４〜ｌＸ１０
９−好ましくはＩ Ｘ １０５〜５ Ｘ １０８個添加
される。mixed. However, in the case of animal cells, IX104 to lX10 for 1+ml of sodium alginate solution
9 - Preferably I x 105 to 5 x 108 are added.
生化学的活性体を添加、もしくは添加しないアルギン酸
ナトリウム溶液は次６１〜３の方法等でゲル化される。The sodium alginate solution with or without the addition of a biochemically active substance is gelated by the following method 61-3.
ゲル化に用いるカルシウムイオンの濃度は０．０１〜１
．０モル濃度程度で、好ましくは０．０２〜０．３モル
濃度程度である。The concentration of calcium ions used for gelation is 0.01-1
．． The concentration is about 0 molar, preferably about 0.02 to 0.3 molar.
１、 注射器やピペット等のノズルからカルシウムイオ
ンを含有する水溶液中に滴下することによりビーズ状の
ゲルが得られる。1. Bead-shaped gel can be obtained by dropping it into an aqueous solution containing calcium ions from a nozzle such as a syringe or pipette.
２、 カルシウムイオンを含有する水溶液中で注射器、
ピペット等のノズルから連続的に吐出させることにより
繊維状のゲルが得られる。2. Syringe in an aqueous solution containing calcium ions,
A fibrous gel can be obtained by continuously discharging it from a nozzle such as a pipette.
３、平板上にキャストするか濾紙もしくはガーゼなどに
含浸させた後、水溶液中のカルシウムイオンと接触させ
ることにより膜状のゲルを得ることができる。3. After casting on a flat plate or impregnating filter paper or gauze, a membrane-like gel can be obtained by contacting with calcium ions in an aqueous solution.
ここに得られたゲルはバリウムイオンを含む溶液に浸漬
したり、該溶液を噴霧したりして処理される。バリウム
イオンの濃度は０．０１〜１．０モル。The gel obtained here is treated by immersing it in a solution containing barium ions or by spraying the solution. The concentration of barium ion is 0.01 to 1.0 mol.
好ましくは０．０２〜０．３モル程度である。処理時間
は １秒〜１時間、ｐＨ３〜１１、温度４〜５０℃程度
である。Preferably it is about 0.02 to 0.3 mol. The treatment time is about 1 second to 1 hour, pH is about 3 to 11, and temperature is about 4 to 50°C.
本発明のバリウムイオンで強化処理されたアルギン酸ゲ
ルは燐酸に対して強い抵抗力をもつようになる。動物細
胞株、ハイブリドーマ等をゲル中に含有させた場合、培
養液にリン酸を含有させ使用するので、特に有効である
。即ち、バリウムイオンで強化処理したものは１週間以
上の長期間リン酸緩衝液に浸漬しても崩壊しないがバリ
ウムイオンで強化処理しないものはリン酸緩衝液に浸漬
して１日以内ですべて崩壊してしまうものである。The barium ion-strengthened alginate gel of the present invention has strong resistance to phosphoric acid. It is particularly effective when animal cell lines, hybridomas, etc. are contained in the gel, since phosphoric acid is used in the culture solution. In other words, those that have been strengthened with barium ions will not disintegrate even if immersed in a phosphate buffer for a long period of one week or more, but those that have not been strengthened with barium ions will completely collapse within one day of being immersed in a phosphate buffer. It's something you end up doing.
次に本発明の実施例を示す。Next, examples of the present invention will be shown.
実施例１゜
２％アルギン酸ナトリウム溶液１００ｍＱ、に、市販の
マウスミエローマ細胞（大日本製薬に、に製）に８リン
パ球を常法により融合したハイブリドーマ培養懸濁液を
１．ＯＸ １０’ケ／ｇゲルになるように添加。Example 1 A hybridoma culture suspension in which 8 lymphocytes were fused to commercially available mouse myeloma cells (manufactured by Dainippon Pharmaceutical Co., Ltd.) using a conventional method was added to 100 mQ of a 2% sodium alginate solution. Add OX at 10'ke/g gel.
混合し、注射器につめ、氷水中の０．１Ｍ塩化カルシウ
ム溶液に滴下し、２０分放置し、粒状化ゲルを得た。The mixture was mixed, filled into a syringe, added dropwise to a 0.1M calcium chloride solution in ice water, and left to stand for 20 minutes to obtain a granulated gel.
さらに粒状化ゲルを４つに分けて、氷水中の０゜１Ｍ塩
化バリウム溶液に０分、５秒、８分、２０分、それぞれ
浸漬した。それぞれを無血清ＲＤＦ培地で２週間３７℃
で培養後、それぞれの１個に０．４ｇの荷重をかけ、収
縮率を測定した。結果は表１に示される。収縮率の算定
は次式による。Furthermore, the granulated gel was divided into four parts and immersed in a 0° 1M barium chloride solution in ice water for 0 minutes, 5 seconds, 8 minutes, and 20 minutes, respectively. Each was incubated at 37°C for 2 weeks in serum-free RDF medium.
After culturing, a load of 0.4 g was applied to each piece, and the shrinkage rate was measured. The results are shown in Table 1. The shrinkage rate is calculated using the following formula.
表１
表１から明らかなように、バリウム処理０秒（対照）即
ち、バリウム処理しない場合は、ゲルは完全につぶれ、
バリウム処理したものは、処理時間が長くなるほど強固
になるのが分かる。Table 1 As is clear from Table 1, when barium treatment was performed for 0 seconds (control), that is, without barium treatment, the gel was completely crushed;
It can be seen that the barium treated material becomes stronger as the treatment time increases.
代理人 弁理士 戸 １）親 男Agent Patent Attorney 1) Parent Male
Claims (1)
宜形状に成型し、次いでバリウムイオンで処理し、強化
することを特徴とする強化されたゲルの製造法。 ２、生化学的活性体を添加したアルギン酸ナトリウム溶
液をカルシウムイオンで適宜形状に成型し、次いでバリ
ウムイオンで処理し、強化することを特徴とする強化さ
れた固定化ゲルの製造法。 ３、アルギン酸ナトリウムがＭ／Ｇ比０．１〜０．８の
アルギン酸ナトリウムである特許請求の範囲第１項又は
第２項記載のゲル又は固定化ゲルの製造法。 ４、生化学的活性体が酵素、微生物、動物細胞又は植物
細胞であることを特徴とする特許請求の範囲第２項記載
の固定化ゲルの製造法。[Claims] 1. A method for producing a reinforced gel, which comprises molding a sodium alginate solution into an appropriate shape with calcium ions, and then treating and strengthening with barium ions. 2. A method for producing a reinforced immobilized gel, which comprises molding a sodium alginate solution containing a biochemically active substance into an appropriate shape with calcium ions, and then treating and strengthening with barium ions. 3. The method for producing a gel or immobilized gel according to claim 1 or 2, wherein the sodium alginate is sodium alginate with an M/G ratio of 0.1 to 0.8. 4. The method for producing an immobilized gel according to claim 2, wherein the biochemically active substance is an enzyme, a microorganism, an animal cell, or a plant cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27169287A JPH066601B2 (en) | 1987-10-29 | 1987-10-29 | Method for producing gel or immobilized gel |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP27169287A JPH066601B2 (en) | 1987-10-29 | 1987-10-29 | Method for producing gel or immobilized gel |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01115901A true JPH01115901A (en) | 1989-05-09 |
| JPH066601B2 JPH066601B2 (en) | 1994-01-26 |
Family
ID=17503514
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP27169287A Expired - Lifetime JPH066601B2 (en) | 1987-10-29 | 1987-10-29 | Method for producing gel or immobilized gel |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH066601B2 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0641196A1 (en) * | 1992-05-29 | 1995-03-08 | Vivorx, Incorporated | Microencapsulation of cells |
| WO2000050103A1 (en) * | 1999-02-25 | 2000-08-31 | Scimed Life Systems, Inc. | Medical devices comprising hydrogel polymers having improved mechanical properties |
| JP2011132395A (en) * | 2009-12-25 | 2011-07-07 | Seiko Epson Corp | Gel manufacturing method |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5718862A (en) * | 1996-04-24 | 1998-02-17 | Hercules Incorporated | Secondary shaping of ionically crosslinked polymer compositions for medical devices |
| WO2021181437A1 (en) * | 2020-03-10 | 2021-09-16 | University Of Petra | A method of preparing alginate micro-particulates |
-
1987
- 1987-10-29 JP JP27169287A patent/JPH066601B2/en not_active Expired - Lifetime
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0641196A1 (en) * | 1992-05-29 | 1995-03-08 | Vivorx, Incorporated | Microencapsulation of cells |
| EP0641196A4 (en) * | 1992-05-29 | 1995-05-03 | Vivorx Inc | Microencapsulation of cells. |
| WO2000050103A1 (en) * | 1999-02-25 | 2000-08-31 | Scimed Life Systems, Inc. | Medical devices comprising hydrogel polymers having improved mechanical properties |
| JP2011132395A (en) * | 2009-12-25 | 2011-07-07 | Seiko Epson Corp | Gel manufacturing method |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH066601B2 (en) | 1994-01-26 |
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