JPH03277967A - Adsorbent for affinity chromatography - Google Patents
Adsorbent for affinity chromatographyInfo
- Publication number
- JPH03277967A JPH03277967A JP2080256A JP8025690A JPH03277967A JP H03277967 A JPH03277967 A JP H03277967A JP 2080256 A JP2080256 A JP 2080256A JP 8025690 A JP8025690 A JP 8025690A JP H03277967 A JPH03277967 A JP H03277967A
- Authority
- JP
- Japan
- Prior art keywords
- adsorbent
- ligand
- carrier
- pectic acid
- gel
- 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.)
- Pending
Links
- 239000003463 adsorbent Substances 0.000 title claims abstract description 27
- 238000001042 affinity chromatography Methods 0.000 title claims abstract description 13
- 239000003446 ligand Substances 0.000 claims abstract description 19
- 229920001277 pectin Polymers 0.000 claims description 11
- 125000006850 spacer group Chemical group 0.000 abstract description 13
- 238000000034 method Methods 0.000 abstract description 11
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 abstract description 5
- 238000010828 elution Methods 0.000 abstract description 3
- 210000002421 cell wall Anatomy 0.000 abstract description 2
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 2
- 239000011159 matrix material Substances 0.000 abstract description 2
- 150000002894 organic compounds Chemical class 0.000 abstract description 2
- 229920002230 Pectic acid Polymers 0.000 abstract 3
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 abstract 3
- 239000010318 polygalacturonic acid Substances 0.000 abstract 3
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 abstract 2
- 239000012620 biological material Substances 0.000 abstract 2
- 229910052799 carbon Inorganic materials 0.000 abstract 1
- 238000001179 sorption measurement Methods 0.000 abstract 1
- 239000007863 gel particle Substances 0.000 description 7
- 239000000243 solution Substances 0.000 description 5
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- UCMIRNVEIXFBKS-UHFFFAOYSA-N beta-alanine Chemical compound NCCC(O)=O UCMIRNVEIXFBKS-UHFFFAOYSA-N 0.000 description 4
- 229950006238 nadide Drugs 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 101710146072 Oxygen-independent coproporphyrinogen III oxidase Proteins 0.000 description 3
- -1 aminoalkyl carboxylic acids Chemical class 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- SLXKOJJOQWFEFD-UHFFFAOYSA-N 6-aminohexanoic acid Chemical compound NCCCCCC(O)=O SLXKOJJOQWFEFD-UHFFFAOYSA-N 0.000 description 2
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229960002684 aminocaproic acid Drugs 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 description 2
- 125000000524 functional group Chemical group 0.000 description 2
- NAQMVNRVTILPCV-UHFFFAOYSA-N hexane-1,6-diamine Chemical compound NCCCCCCN NAQMVNRVTILPCV-UHFFFAOYSA-N 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 239000013076 target substance Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 1
- UXMZXDQTOZIONB-UHFFFAOYSA-N 6-aminooctanoic acid Chemical compound CCC(N)CCCCC(O)=O UXMZXDQTOZIONB-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 1
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- GKXVJHDEWHKBFH-UHFFFAOYSA-N [2-(aminomethyl)phenyl]methanamine Chemical compound NCC1=CC=CC=C1CN GKXVJHDEWHKBFH-UHFFFAOYSA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- LSTJLLHJASXKIV-UHFFFAOYSA-N amino hexanoate Chemical compound CCCCCC(=O)ON LSTJLLHJASXKIV-UHFFFAOYSA-N 0.000 description 1
- 229940124277 aminobutyric acid Drugs 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 229940000635 beta-alanine Drugs 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000012050 conventional carrier Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 150000004985 diamines Chemical class 0.000 description 1
- QFTYSVGGYOXFRQ-UHFFFAOYSA-N dodecane-1,12-diamine Chemical compound NCCCCCCCCCCCCN QFTYSVGGYOXFRQ-UHFFFAOYSA-N 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 239000013077 target material Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明は、アフィニティクロマトグラフィー用吸着体に
関する。DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an adsorbent for affinity chromatography.
特定の蛋白質・ウィルス等を精製収得するには従来遠心
分離法・イオン交換法・ゲル濾過法等が採用されており
、これらの方法を適宜組み合わせたり、反復して用いら
れる。ところが、これらの方法を組み合わせ、あるいは
反復して実施する場合、各工程毎に複雑な操作を必要と
し、また、長時間要する上それらの工程を経るに従って
目的物の損失減量が生じる問題がある。Conventionally, centrifugation, ion exchange, gel filtration, and the like have been used to purify and obtain specific proteins, viruses, etc., and these methods are appropriately combined or used repeatedly. However, when these methods are combined or repeated, there are problems in that each step requires complicated operations, takes a long time, and loses and reduces the amount of the target material as the steps go through them.
別の精製法としてアフィニテイクロマトグラフィーがあ
る。このものは、生化学的な特異性のあるアフィニティ
（！ｌ！和力）を相互に持つ２種類の物質または物質
群の一方をリガンドとして水不溶性の担体に固定化して
固定相とし、この固定相に対するアフィニテイの差を利
用して目的物質をこれに混在する不純物から分離・精製
するクロマトグラフィーであって、生化学、特に酵素化
学の分野に於ける９踏・精製に広く用いられている。上
記担体としては、従来、デキストラン・アガロ−弱いた
め高流速が得られず、また価格的に非常に高価なもので
あるため、工業スケールでは、導入し難いという問題が
発生している。Another purification method is affinity chromatography. This is a stationary phase in which one of two substances or a group of substances that have a biochemically specific affinity (!l! power) for each other is immobilized as a ligand on a water-insoluble carrier. Chromatography is a method of separating and purifying a target substance from impurities mixed therein by making use of differences in affinity between phases, and is widely used for steps and purification in the field of biochemistry, especially enzyme chemistry. Conventionally, the above-mentioned carrier has been dextran agaro, which is weak and cannot obtain a high flow rate, and is very expensive, so it is difficult to introduce it on an industrial scale.
（発明の目的）
本発明は、従来のアフィニティクロマトグラフィーに於
ける問題、特に吸着体に於ける問題を解決するためにな
されたものであって、物理的強度に優れているのみなら
ず、吸着及び溶出効率が高く、しかも目的物質以外の不
純物の非特異的な吸着がない価格的に安価なアフィニテ
ィクロマトグラフィー用吸着体を提供することを目的と
する。(Purpose of the Invention) The present invention was made to solve problems in conventional affinity chromatography, particularly in adsorbents. Another object of the present invention is to provide an inexpensive adsorbent for affinity chromatography that has high elution efficiency and does not non-specifically adsorb impurities other than the target substance.
本発明によるアフィニティクロマトグラフィー用吸着体
は、ペクチン酸−セルロースゲルを担体とし、この担体
上にリガンドな共有結合にて固定化されていることを特
徴とする。必要に応じてリガンドの吸着体上での自由度
を高めるために、吸着体とリガンドとをスペーサ基を介
在させて共有結合にて結合させることができる。このス
ペーサ基は、予め吸着体に結合させておき、この後に、
このスペーサ基にリガンドを結合させてもよく、あるい
は、スペーサ基を予めリガンドに結合させ、これを吸着
体に結合させてもよい。さらに必要に応じて吸着体及び
リガンドの両者に予めスペーサ基を結合させ、これらを
相互に結合さすこともできる。The adsorbent for affinity chromatography according to the present invention is characterized in that it uses a pectic acid-cellulose gel as a carrier, and is immobilized on the carrier by a covalent bond such as a ligand. If necessary, in order to increase the degree of freedom of the ligand on the adsorbent, the adsorbent and the ligand can be bonded to each other through a covalent bond with a spacer group interposed therebetween. This spacer group is bonded to the adsorbent in advance, and then,
A ligand may be bonded to this spacer group, or the spacer group may be bonded to a ligand in advance and this may be bonded to the adsorbent. Furthermore, if necessary, spacer groups can be bonded to both the adsorbent and the ligand in advance to bond them to each other.
上記スペーサ基として用い得る化合物は少なくとも二官
能性の有機化合物であり、多官能性の重合体を排除する
ものではないが、特に、炭素数１〜１２の炭素鎖基を有
する二官能性の有機化合物が好ましい、このようなスペ
ーサ基として機能する化合物の具体例としてたとえば、
ヘキサメチレンジアミン・ドデカメチレンジアミン・キ
シリレンジアミン等のジアミン類、グリシン・β−アミ
ノプロピオン酸・Ｔ−アミノ酪酸・ε−アミノカプロン
酸・ε−アミノカプリル酸等のアミノアルキルカルボン
酸、リジン・グルタミン酸・β−アラニン・アルギニン
等のアミノ酸類等が好ましく用いられるが、これらに限
定されるものではない。The compound that can be used as the spacer group is at least a difunctional organic compound, and does not exclude polyfunctional polymers. Specific examples of compounds that function as such spacer groups, which are preferably compounds, include, for example,
Diamines such as hexamethylene diamine, dodecamethylene diamine, xylylene diamine, aminoalkyl carboxylic acids such as glycine, β-aminopropionic acid, T-aminobutyric acid, ε-aminocaproic acid, ε-aminocaprylic acid, lysine, glutamic acid, etc. Amino acids such as β-alanine and arginine are preferably used, but are not limited to these.
前記したペクチン酸−セルロースゲル粒子に、直接にリ
ガンドを共有結合にて結合するか、または、ペクチン酸
−セルロースゲル粒子にスペーサ基を結合し、このスペ
ーサ基にリガンドを共有結合にて結合するための方法は
、特に制限されず、従来より知られている任意の方法に
よる事ができる。例えば、好ましい方法の一つとして、
結合試薬として水溶性カルボジイミドを用いる方法を挙
げることができる。例えばアミノアルキルカルボン酸を
、ペクチン酸−セルロースゲル粒子に結合サセ、次いで
、このペクチン酸−セルロースゲル粒子に結合されたア
ミノアルキルカルボン酸に水溶性カルボジイミドを用い
て、同様にしてリガンドを共有結合にて結合させること
ができる。To bond a ligand directly to the above-mentioned pectic acid-cellulose gel particles by a covalent bond, or to bond a spacer group to the pectic acid-cellulose gel particles and bond the ligand to this spacer group by a covalent bond. The method is not particularly limited, and any conventionally known method can be used. For example, one of the preferred methods is
A method using water-soluble carbodiimide as a binding reagent can be mentioned. For example, an aminoalkyl carboxylic acid is bound to pectic acid-cellulose gel particles, and then a water-soluble carbodiimide is used to bind the aminoalkyl carboxylic acid bound to the pectic acid-cellulose gel particles in a similar manner to covalently bond the ligand. can be combined.
係る方法に於いて用いる水溶性カルボジイミドとしては
、例えば、ｌ−エチル−３−（３−ジメチルアミノプロ
ピル）カルボジイミド塩酸塩・ｌシクロへキシル−３−
（２−モノホリノエチル）カルボジイミド−メトーｐ−
トルエンスルホネーをペクチン酸−セルロースゲル粒子
に結合させるには、従来より知られている通常の方法及
び条件によることができる。Examples of the water-soluble carbodiimide used in such a method include 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride and 1-cyclohexyl-3-
(2-monopholinoethyl)carbodiimide-metho p-
Toluenesulfone can be bound to the pectic acid-cellulose gel particles by conventional methods and conditions known in the art.
また官能基が水酸基である時は、臭化シアンにより反応
させ、官能基を活性化することによってスペーサ基を結
合させ、次いで上記と同様にしてリガンドを共有結合に
て結合させることができる。When the functional group is a hydroxyl group, a spacer group can be bound by activating the functional group by reaction with cyanogen bromide, and then a ligand can be bound by a covalent bond in the same manner as above.
また、ペクチン酸−セルロースゲル粒子に直接にリガン
ドを結合させることもできる。Alternatively, a ligand can be directly bound to the pectic acid-cellulose gel particles.
るアフィニティクロマトグラフィー用吸着体を分Ｍ・精
製することができる。The adsorbent for affinity chromatography can be purified in minutes.
本発明によるアフィニティクロマトグラフィー用吸着体
は、従来のアフィニティクロマトグラフィーにおける膨
潤ゲルと同様にして用いることができる。従って吸着体
は、カラム法・回分法の両者に使用し得る。The adsorbent for affinity chromatography according to the present invention can be used in the same manner as a swelling gel in conventional affinity chromatography. Therefore, the adsorbent can be used in both column and batch methods.
〔実施例）
以下に実施例を挙げて本発明を説明するが、本発明は、
これら実施例により何ら限定されるものではない。[Example] The present invention will be described below with reference to Examples.
The present invention is not limited in any way by these Examples.
実施例１
＋１１ ニコチンアミドアデニンジヌクレオチド（以
下ＮＡＤと略す）吸着体の調製
ペクチン酸−セルロースゲル５０ｍＪ（乾燥重量８ｇ）
を４０ｇのＣＮＢ ｒでｐＨ１１で活性化し、洗浄後、
これに８−アミノカプロン酸の溶液（２０ｇを２００ｍ
ｊ！の０． Ｉ Ｍ Ｎ ａ ＨＣＯｓＯ，０１Ｍ
ＨＣｊ・０．５Ｍ ＮａＣ１溶液で順次洗浄し、最
後に十分水洗した。洗浄したゲルを次いでジシクロへキ
シルカルボジイミドのピリジン溶液（８０ｇ／ １９２
ｍＪ）を加え、密栓してｌＯ日間室温で撹拌する。その
後ガラスフィルターで濾取し水、エタノールで洗浄しＮ
ＡＤ吸着体を得た。Example 1 +11 Preparation of nicotinamide adenine dinucleotide (hereinafter abbreviated as NAD) adsorbent Pectic acid-cellulose gel 50 mJ (dry weight 8 g)
was activated with 40 g of CNBr at pH 11, and after washing,
Add a solution of 8-aminocaproic acid (20g to 200ml
j! 0. I M N a HCOsO, 01M
It was washed successively with HCj/0.5M NaCl solution, and finally thoroughly washed with water. The washed gel was then treated with a solution of dicyclohexylcarbodiimide in pyridine (80 g/192
mJ), capped tightly and stirred at room temperature for 10 days. After that, it was filtered through a glass filter and washed with water and ethanol.
An AD adsorbent was obtained.
（比較例１）
比較のために、セファロース４Ｂ（ファルマシア社製ア
ガロース系担体）２００ｍｊ （乾燥重量８ｇ）を４０
ｇＣＮＢｒでｐＨ１ｌで活性化し、洗浄後これにε−ア
ミノカプロン酸の溶液を加え、上記（実施例１）と同様
に反応を行い、次いでＮＡＤｔｇ液（１，６ｇを水４８
ｍｌ溶解）ジシクロへキシルカルボジイミドのピリジン
溶液を加えＮＡＤ吸着体を調製した。その結果を図１に
示す。(Comparative Example 1) For comparison, 200 mj of Sepharose 4B (agarose carrier manufactured by Pharmacia) (dry weight 8 g) was
After activation with gCNBr at pH 1l, a solution of ε-aminocaproic acid was added to this and the reaction was carried out in the same manner as described above (Example 1).
A NAD adsorbent was prepared by adding a pyridine solution of dicyclohexylcarbodiimide (dissolved in ml). The results are shown in Figure 1.
（２） グリセルアルデヒド３リン酸脱水素酵素・乳
酸脱水素酵素・牛血清アルブミン（以下ＣＰＤＨ・ＬＤ
Ｈ−ＢＳＡと略す）混合溶液からの各成分の分離
上で得たＮＡＤ吸着体を１０１０Ｘ１０３のカラムに充
填し試料： ＣＰＤＨ（４，Ｉ Ｕ） ・Ｌ Ｄ Ｈ
次ぎに０．１５ｍＭ ＮＡＤｌｌ−１０ｍＭ ＮＡ
ＤＨで順次溶出を行った。なお流速は、ｌ ｒｎ １
／ ８ｍ１ｎとした。得られたカラムクロマトの結果を
図１に示す。(2) Glyceraldehyde 3-phosphate dehydrogenase, lactate dehydrogenase, bovine serum albumin (CPDH, LD)
The NAD adsorbent obtained by separating each component from the mixed solution (abbreviated as H-BSA) was packed into a 1010 x 103 column, and sample: CPDH (4, I U) ・L D H
Then 0.15mM NADll-10mM NA
Elution was performed sequentially with DH. Note that the flow rate is l rn 1
/8m1n. The results of the column chromatography obtained are shown in FIG.
本発明の吸着体によれば、比較例のそれに比べＣＰＤＨ
−ＬＤＨ−ＢＳＡの回収効率が高く、また流速を上げて
も回収効率に余り影響がないことが明らかである。（表
１）
〔発明の効果〕
以上のように、本発明によるアフイニテイクロマトグラ
フィー用吸着体は、ペクチン酸−セルロースゲル粒子を
担体とし、この担体上に、リガンドが共有結合にて結合
されている。従って係る吸着体は、生体物質移動の場で
ある細胞壁マトリックスを保持したペクチン酸−セルロ
ースゲルを担体としているため、生体物質に対する安定
性に優れている。また、硬質ゲルであるため、従来高流
速を得るためには、ビーズ状ゲルでなければならなかっ
たが本担体は、破砕型にもかかわらず従来担体が非常に
安価に提供され得るため、当然のごとく、本担体を用い
たアフィニティクロマトグラフィー用吸着体も安価とな
り、本吸着体を工業スケールに導入した場合大きなメリ
ットを与え得るものである。According to the adsorbent of the present invention, compared to that of the comparative example, CPDH
It is clear that the recovery efficiency of -LDH-BSA is high and that increasing the flow rate does not significantly affect the recovery efficiency. (Table 1) [Effects of the Invention] As described above, the adsorbent for Affinity chromatography according to the present invention uses pectic acid-cellulose gel particles as a carrier, and a ligand is bound to the carrier by a covalent bond. ing. Therefore, such an adsorbent has excellent stability against biological substances because it uses a pectic acid-cellulose gel as a carrier that retains a cell wall matrix, which is a site for the transfer of biological substances. In addition, because it is a hard gel, conventionally it had to be a bead-like gel in order to obtain a high flow rate, but this carrier is a crushed type, but since conventional carriers can be provided at a very low cost, it is natural that As shown, the adsorbent for affinity chromatography using this carrier is also inexpensive, and it can bring great benefits when introduced on an industrial scale.
表table
Claims (1)
上にリガンドが共有結合にて固定化されていることを特
徴とするアフィニティクロマトグラフィー用吸着体。1. An adsorbent for affinity chromatography, comprising a pectic acid-cellulose gel as a carrier, on which a ligand is covalently immobilized.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2080256A JPH03277967A (en) | 1990-03-27 | 1990-03-27 | Adsorbent for affinity chromatography |
| PCT/JP1990/001493 WO1991007427A1 (en) | 1989-11-16 | 1990-11-15 | Carrier for column chromatography, process for separating and purifying water-soluble polymeric substance using said carrier, novel pectic acid-cellulose gel and process for preparing the same, and adsorbent for affinity chromatography |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2080256A JPH03277967A (en) | 1990-03-27 | 1990-03-27 | Adsorbent for affinity chromatography |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH03277967A true JPH03277967A (en) | 1991-12-09 |
Family
ID=13713234
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2080256A Pending JPH03277967A (en) | 1989-11-16 | 1990-03-27 | Adsorbent for affinity chromatography |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH03277967A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010095673A1 (en) * | 2009-02-20 | 2010-08-26 | チッソ株式会社 | Cellulose gel for purification of immunoglobulin |
-
1990
- 1990-03-27 JP JP2080256A patent/JPH03277967A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010095673A1 (en) * | 2009-02-20 | 2010-08-26 | チッソ株式会社 | Cellulose gel for purification of immunoglobulin |
| CN102326075A (en) * | 2009-02-20 | 2012-01-18 | Jnc株式会社 | Cellulose gel for purification of immunoglobulin |
| US8912117B2 (en) | 2009-02-20 | 2014-12-16 | Jnc Corporation | Cellulose gel for purification of immunoglobulin |
| JP5692059B2 (en) * | 2009-02-20 | 2015-04-01 | Jnc株式会社 | Cellulosic gel for immunoglobulin purification |
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