WO1996008716A1 - Capillary affinity electrophoresis, affinity matrix for implementing said electrophoresis and method of manufacture - Google Patents
Capillary affinity electrophoresis, affinity matrix for implementing said electrophoresis and method of manufacture Download PDFInfo
- Publication number
- WO1996008716A1 WO1996008716A1 PCT/FR1995/001176 FR9501176W WO9608716A1 WO 1996008716 A1 WO1996008716 A1 WO 1996008716A1 FR 9501176 W FR9501176 W FR 9501176W WO 9608716 A1 WO9608716 A1 WO 9608716A1
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- WIPO (PCT)
- Prior art keywords
- affinity
- electrophoresis
- capillary
- peg
- soluble polymer
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44747—Composition of gel or of carrier mixture
Definitions
- the present invention relates to an improvement relating to an analysis technique called capillary affinity electrophoresis, also called high performance capillary electrophoresis (ECHP), and an affinity support for the implementation of this technique and to a method for the manufacture of this support.
- capillary affinity electrophoresis also called high performance capillary electrophoresis (ECHP)
- ECHP high performance capillary electrophoresis
- This analysis technique is currently carried out using a device which consists of a silica capillary, which has an internal diameter of between 25 and 100 ⁇ m, and each end of which is immersed in containers filled with buffers. A voltage of the order of 20 to 30 V is applied via two electrodes immersed in these containers.
- electrophoresis a person skilled in the art knows that high electric fields allow rapid separations.
- capillary electrophoresis where the injection and the detection volume are minimal, the only factor of widening of the peaks is due to the axial diffusion. The greater the speed of migration of chemical species outside the capillary, the more the peaks are resolved. This electric speed is obtained for electric fields of the order of 150 to 300 V / cm.
- capillary affinity electrophoresis a person skilled in the art proposes to fill the capillary with an agarose gel coupled with a ligand or to carry out a derivatization of the wall of the capillary and to couple the ligand.
- these known techniques of capillary affinity electrophoresis are difficult to use with all types of bio-specific ligands, specific or pseudo-bio-specific groups, including metal chelates such as copper (II) imi ⁇ odiacetate. or IDA-Cu (II)
- this current technique has the drawback that the ligand remains fixed in the capillary: the regeneration of the latter after each separation can turn out to be very difficult or even impossible. This is particularly the case for metal chelates because a release of the metal (due to the electric field or a great affinity for the protein used) cannot be compensated for and the properties of the capillary will therefore be modified.
- One of the aims of the present invention is therefore to provide a technique of affinity capillary electrophoresis which makes it possible to overcome these drawbacks.
- Another object of the present invention is to provide such an electrophoresis technique allowing preparation, regeneration and satisfactory stability of the capillary.
- the present invention also aims to provide an affinity support for the implementation of this capillary affinity electrophoresis as well as a product for the manufacture of this support.
- This affinity support is constituted by an affinity ligand coupled to a covalently soluble polymer.
- the soluble polymer is an organic aliphatic compound, with a linear or cyclic chain, unbranched, biocompatible and having a mass greater than 4,996.10 "24 kg (3,000 dates).
- the viscosity of this soluble polymer is less than 5 mPa.s.
- the ultraviolet is weakly absorbed by this polymer, which makes it possible to detect proteins at wavelengths between 214 and 280 nm.
- the soluble polymer is preferably polyethylene glycol (or PEG) or one of its derivatives, such as methoxy-polyethylene glycol (or m. PEG).
- PEG polyethylene glycol
- the PEG has a mass of the order of 8,302.10 "2 * kg. (5,000 daltons) and is linear and unbranched. The viscosity is less than 5 mPa.s. It is soluble in water, the benzene, acetonitrile and, when hot, in acetone, dioxane, toluene, for example.
- Various additives can be used, up to one mole in 5% of PEG: either salts such as sodium (NaCI), potassium sulphate (r ⁇ SC ⁇ , or glycine-betaine, these additives being mentioned without limitation.
- This polymer is coupled to an affinity ligand such as, for example, lectins, immobilized metal ions.
- the affinity support thus obtained is in liquid form and constitutes the buffer filling the containers of the electrophoresis device.
- Example 1 PEG coupled to a lectin
- Methoxy-PEG-Succinyl -ConA is used as affinity support in order to separate RNases A and B from bovine pancreas. These two RNases have identical sequences in animated acids, but RNase B is gtycolysed in Asn 34 while RNase A is not glycolysed.
- Table 2 are grouped the results of capillary affinity electrophoresis carried out, one without an affinity ligand, the other with the above affinity ligand.
- Example 2 PEG coupled to an immobilized metal ion
- the methoxy-PEG 5000 IDA-Cu (II) is used as the affinity support and the methoxy PEG 5000 is used as the reference support.
- the cytochromes c of tuna heart, horse heart and Candida Krusei are to be separated: these different species of cytochromic c are proteins which have substantially the same molecular weight and the same isoelectric point. We know that they differ, however, in their numbers of histidine residues accessible on the surface. In Table 3 below, the migration times of each cytochrome c are grouped according to the support used.
Abstract
Capillary affinity electrophoresis characterized by having, as the affinity carrier, an affinity ligand coupled to a soluble polymer such as polyethyleneglycol (PEG) or a derivative thereof.
Description
Titre : Electrophorèse capillaire d'affinité, matrice d'affinité pour la mise en oeuvre de cette électrophorèse et son procédé de fabrication.Title: Capillary affinity electrophoresis, affinity matrix for the implementation of this electrophoresis and its manufacturing process.
La présente invention concerne un perfectionnement relatif à une technique d'analyse dite électrophorèse capillaire d'affinité, également dénommée électrophorèse capillaire haute performance (ECHP), et un support d'affinité pour la mise en oeuvre de cette technique et à un procédé pour la fabrication de ce support.The present invention relates to an improvement relating to an analysis technique called capillary affinity electrophoresis, also called high performance capillary electrophoresis (ECHP), and an affinity support for the implementation of this technique and to a method for the manufacture of this support.
Cette technique d'analyse est actuellement réalisée au moyen d'un appareil qui consiste en un capillaire de silice, qui a un diamètre intérieur compris entre 25 et 100 μm, et dont chaque extrémité plonge dans des récipients rempli de tampons. Un voltage de l'ordre de 20 à 30 V est appliqué par l'intermédiaire de deux électrodes plongeant dans ces récipients. En électrophorèse, l'homme du métier sait que des champs électriques élevés permettent des séparations ra¬ pides. Dans le cas de l'électrophorèse capillaire où l'injection et le volume de dé¬ tection sont minimaux, le seul facteur d'élargissement des pics est dû à la diffusion axiale. Plus la rapidité de la migration des espèces chimiques à l'extérieur du capil- laire est importante, plus les pics sont résolus. Cette vitesse électrique est obtenue pour des champs électriques de l'ordre de 150 à 300 V/cm.This analysis technique is currently carried out using a device which consists of a silica capillary, which has an internal diameter of between 25 and 100 μm, and each end of which is immersed in containers filled with buffers. A voltage of the order of 20 to 30 V is applied via two electrodes immersed in these containers. In electrophoresis, a person skilled in the art knows that high electric fields allow rapid separations. In the case of capillary electrophoresis where the injection and the detection volume are minimal, the only factor of widening of the peaks is due to the axial diffusion. The greater the speed of migration of chemical species outside the capillary, the more the peaks are resolved. This electric speed is obtained for electric fields of the order of 150 to 300 V / cm.
Dans la technique d'électrophorèse capillaire d'affinité, l'homme du métier propose de remplir le capillaire d'un gel d'agarose couplé avec un ligand ou de réaliser une dérivatisation de la paroi du capillaire et d coupler le ligand. Mais ces techniques connues d'électrophorèse capillaire d'affinité sont difficiles à mettre en oeuvre avec tous types de ligands bio-spécifiques, groupes spécifiques ou pseudo-bio-spécifiques, y compris les chélates métalliques tels que l'imiπodiacétate de cuivre (II) ou IDA-Cu (II)In the technique of capillary affinity electrophoresis, a person skilled in the art proposes to fill the capillary with an agarose gel coupled with a ligand or to carry out a derivatization of the wall of the capillary and to couple the ligand. However, these known techniques of capillary affinity electrophoresis are difficult to use with all types of bio-specific ligands, specific or pseudo-bio-specific groups, including metal chelates such as copper (II) imiπodiacetate. or IDA-Cu (II)
En particulier, cette technique actuelle a pour inconvénient le fait que le ligand reste fixé dans le capillaire : la régénération de ce dernier après chaque sé¬ paration peut se révéler être très difficile voire même impossible. Ceci est tout par¬ ticulièrement le cas pour les chélates métalliques car un relargage du métal (du fait du champ électrique ou d'une grande affinité pour la protéine utilisée) ne pourra être compensée et les propriétés du capillaire seront par conséquent modifiées. Aussi un des buts de la présente invention est-il de fournir une tech¬ nique d'électrophorèse capillaire d'affinité qui permet de pallier ces inconvénients.In particular, this current technique has the drawback that the ligand remains fixed in the capillary: the regeneration of the latter after each separation can turn out to be very difficult or even impossible. This is particularly the case for metal chelates because a release of the metal (due to the electric field or a great affinity for the protein used) cannot be compensated for and the properties of the capillary will therefore be modified. One of the aims of the present invention is therefore to provide a technique of affinity capillary electrophoresis which makes it possible to overcome these drawbacks.
Un autre but de la présente invention est de fournir une telle technique d'électrophorèse permettant une préparation, une régénération et une stabilité satisfaisante du capillaire.
Ces buts, ainsi que d'autres qui apparaîtront par la suite, sont atteints par une électrophorèse capillaire d'affinité qui est caractérisée, selon la présente invention, par le fait qu'on utilise comme support d'affinité un ligand d'affinité couplé à un polymère soluble. Avantageusement, le polymère soluble est le polyéthylène glycol (PEG) ou un de ses dérivés.Another object of the present invention is to provide such an electrophoresis technique allowing preparation, regeneration and satisfactory stability of the capillary. These aims, as well as others which will appear subsequently, are achieved by capillary affinity electrophoresis which is characterized, according to the present invention, by the fact that a coupled affinity ligand is used as the affinity support. to a soluble polymer. Advantageously, the soluble polymer is polyethylene glycol (PEG) or one of its derivatives.
La présente invention a également pour but de fournir un support d'affinité pour la mise en oeuvre de cette électrophorèse capillaire d'affinité ainsi qu'un produit pour la fabrication de ce support Ce support d'affinité est constituée par un ligand d'affinité couplé à un polymère soluble par covalence.The present invention also aims to provide an affinity support for the implementation of this capillary affinity electrophoresis as well as a product for the manufacture of this support. This affinity support is constituted by an affinity ligand coupled to a covalently soluble polymer.
Selon la présente invention; le polymère soluble est un composé orga¬ nique aliphatique, à chaîne linéaire ou cyclique, non ramifié, biocompatible et ayant une masse supérieure à 4.996.10"24 kg (3.000 datons). La viscosité de ce poly- mère soluble est inférieure à 5 mPa.s. Par ailleurs, l'ultra-violet est faiblement ab¬ sorbé par ce polymère, ce qui permet une détection des protéines à des longueurs d'onde comprises entre 214 et 280 nm.According to the present invention; the soluble polymer is an organic aliphatic compound, with a linear or cyclic chain, unbranched, biocompatible and having a mass greater than 4,996.10 "24 kg (3,000 dates). The viscosity of this soluble polymer is less than 5 mPa.s. Furthermore, the ultraviolet is weakly absorbed by this polymer, which makes it possible to detect proteins at wavelengths between 214 and 280 nm.
Le polymère soluble est de préférence le polyéthylène glycol (ou PEG) ou un de ses dérivés, tel que le méthoxy-polyéthylène-glycol (ou m. PEG). Le PEG présente une masse de l'ordre de 8,302.10"2* kg. (5.000 dal- tons) et est linéaire et non ramifié. La viscosité est inférieure à 5 mPa.s. Il est so¬ luble dans l'eau, le benzène, l'acétonitrile et, à chaud, dans l'acétone, le dioxane, le toluène, par exemple. On peut utiliser, jusqu'à une mole dans 5 % de PEG, différents additifs : soit des sels tels que le chlorure de sodium (NaCI), le sulfate de potasium (r^SCλ , soit la glycine-bétaïne, ces additifs étant cités de façon non limitative.The soluble polymer is preferably polyethylene glycol (or PEG) or one of its derivatives, such as methoxy-polyethylene glycol (or m. PEG). The PEG has a mass of the order of 8,302.10 "2 * kg. (5,000 daltons) and is linear and unbranched. The viscosity is less than 5 mPa.s. It is soluble in water, the benzene, acetonitrile and, when hot, in acetone, dioxane, toluene, for example. Various additives can be used, up to one mole in 5% of PEG: either salts such as sodium (NaCI), potassium sulphate (r ^ SCλ, or glycine-betaine, these additives being mentioned without limitation.
Le couplage de différents ligands au PEG ou à des dérivés de celui-ci a été réalisé. Ceci est regroupé dans le tableau 1 ci-après.
Tableau 1The coupling of different ligands to PEG or to derivatives thereof has been carried out. This is grouped in Table 1 below. Table 1
Dérivé du PEG Group* fonctionnel Réaction de couplage de ligandDerivative of the functional PEG Group * Ligand coupling reaction
R a iné secondaire ouSecondary or
PEG-CI HN Direct R ternairePEG-CI HN Direct R ternary
PEG-NHj R-COOH DCC, cataiyse possible par HOBT dans des Suivants organiques EDC, possible via NHS-ester dans H OPEG-NHj R-COOH DCC, cataiysis possible by HOBT in organic EDC Following, possible via NHS-ester in H O
R-CHO Base de Schiff , réduite par Na BU* (sucres, nudéαttdes)R-CHO Schiff base, reduced by Na BU * (sugars, nudéαttdes)
00
R-CHO pKa = 3, réaction possible en condrttons (sucres, nudéotirjes) acidesR-CHO pKa = 3, possible reaction in acid condrttons (sugars, nudeotirjes)
PEG-O-CHT-C-NH-NH, (Hydrazide)PEG-O-CHT-C-NH-NH, (Hydrazide)
00
R-NH] Catalyse par aminé, ether ou thioétnerR-NH] Amine, ether or thioether catalysis
PEG-O-OHrCH-CHj R-OH (Epoxydβ) R-SHPEG-O-OHrCH-CHj R-OH (Epoxydβ) R-SH
PE&O-OHrCH-CHj-CHO R-NH2 Base de Schiff, réduite par NcBHa (Aldéhyde)PE & O-OHrCH-CHj-CHO R-NH 2 Schiff base, reduced by NcBHa (Aldehyde)
PEG-O-Succ R-NH2 DCC, possible HOBT- cataiyse possible par HOBTdans un solvant organiquePEG-O-Succ R-NH 2 DCC, possible HOBT- cataiysis possible by HOBT in an organic solvent
EDC, possible via NHS-Ester dans H7O
= rv s ca oxy sEDC, possible via NHS-Ester in H 7 O = rv s ca oxy s
On couple à ce polymère un ligand d'affinité tel que, par exemple, des lectines, des ions métalliques immobilisés. Le support d'affinité ainsi obtenu se présente sous forme liquide et constitue le tampon remplissant les récipients de l'appareil d'électrophorèse.This polymer is coupled to an affinity ligand such as, for example, lectins, immobilized metal ions. The affinity support thus obtained is in liquid form and constitutes the buffer filling the containers of the electrophoresis device.
Deux exemples seront ci-après donnés à titre non limitatif, pour mieux faire comprendre les avantages de la présente invention.Two examples will be given below without implied limitation, to better understand the advantages of the present invention.
Exemple 1 : PEG couplé à une lectineExample 1: PEG coupled to a lectin
On utilise comme support d'affinité du méthoxy-PEG-Succinyl -ConA afin de séparer les RNases A et B de pancréas bovin. Ces deux RNases ont des séquences identiques en acides animés, mais la RNase B est gtycolysée en Asn 34 alors que la RNase A n'est pas glycolysée.
Dans le tableau 2 ci-après sont regroupés les résultats d'électrophorèse capillaire d'affinité réalisée l'une sans ligand d'affinité, l'autre avec le ligand d'afinité ci-dessus. Tableau 2: Temps de migration des différentes protéines étudiées. to = temps de migration sans le ligand mPEG-Con A dans le méthoxy - PEG 5000 t = temps de migration avec le ligand mPEG-Con A dans le méthoxy - PEG 5000- IDA-Cu (II)Methoxy-PEG-Succinyl -ConA is used as affinity support in order to separate RNases A and B from bovine pancreas. These two RNases have identical sequences in animated acids, but RNase B is gtycolysed in Asn 34 while RNase A is not glycolysed. In Table 2 below are grouped the results of capillary affinity electrophoresis carried out, one without an affinity ligand, the other with the above affinity ligand. Table 2: Migration times of the different proteins studied. to = migration time without the mPEG-Con A ligand in methoxy - PEG 5000 t = migration time with the mPEG-Con A ligand in methoxy - PEG 5000- IDA-Cu (II)
Protéine to(miπ) t(min) t-to (min)Protein to (miπ) t (min) t-to (min)
RNase A 9,9 10,2 0,3RNase A 9.9 10.2 0.3
RNase B 12,5 15,7 3,2RNase B 12.5 15.7 3.2
II apparaît que les temps de migration sont bien distincts, ce qui se traduit lors de la mesure par l'obtention de pics bien identifiables. Exemplaire 2 : PEG couplé à un ion métallique immobiliséIt appears that the migration times are very distinct, which results in the measurement by obtaining clearly identifiable peaks. Example 2: PEG coupled to an immobilized metal ion
On utilise comme support d'affinité le méthoxy-PEG 5000 IDA-Cu (II) et comme support de référence le méthoxy PEG 5000. Par électrophorèse on veut séparer les cytochromes c de coeur de thon, de coeur de cheval et de Candida Krusei : ces différentes espèces de cyto- chrome c sont des protéines qui ont sensiblement le même poids moléculaire et le même point isoélectrique. On sait qu'ils diffèrent, néanmoins, de par leurs nombres de résidus histidine accessibles en surface. Dans le tableau 3 ci-après, sont regroupés les temps de migration de chaque cytochrome c en fonction du support utilisé.The methoxy-PEG 5000 IDA-Cu (II) is used as the affinity support and the methoxy PEG 5000 is used as the reference support. By electrophoresis, the cytochromes c of tuna heart, horse heart and Candida Krusei are to be separated: these different species of cytochromic c are proteins which have substantially the same molecular weight and the same isoelectric point. We know that they differ, however, in their numbers of histidine residues accessible on the surface. In Table 3 below, the migration times of each cytochrome c are grouped according to the support used.
Tableau 3 : Temps de migration des différents cytochromes étudiés to = temps de migration dans le méthoxy-PEG 5000 t = temps de migration dans le méthoxy-PEG 5000-IDA-Cu (II)Table 3: Migration times of the various cytochromes studied to = migration times in methoxy-PEG 5000 t = migration times in methoxy-PEG 5000-IDA-Cu (II)
Protéine to(min) t(min)a t-to (min)Protein to (min) t (min) a t-to (min)
Cyt c de coeur de thon 12,6 12,6 0 Cyt c de coeur de cheval 12,2 13,2 1,0 Cyt c Candida Krusei 17,3 20,5 3,2
a) Les valeurs sont corrigées en fonction des différences de flux électroosmotiques pour les deux supports polymères, avec ou sans ligand.Cyt c of tuna heart 12.6 12.6 0 Cyt c of horse heart 12.2 13.2 1.0 Cyt c Candida Krusei 17.3 20.5 3.2 a) The values are corrected as a function of the differences in electroosmotic fluxes for the two polymer supports, with or without ligand.
I! apparaît que la présence du ligand couplé permet une bonne identifi¬ cation des protéines de structure très voisine, leur caractérisation et, par suite, leur séparation.
I! It appears that the presence of the coupled ligand allows good identifi¬ cation of proteins of very similar structure, their characterization and, consequently, their separation.
Claims
1. Electrophorèse capillaire d'affinité, caractérisée par le fait qu'on uti¬ lise comme supportd'affinité un ligand d'affinité couplé à un polymère soluble.1. Capillary affinity electrophoresis, characterized in that an affinity ligand coupled to a soluble polymer is used as the affinity support.
55
2. Electrophorèse capillaire d'affinité selon la revendication 1 , caracté¬ risée par le fait que le polymère soluble est du polyéthylène glycol (PEG) ou un de ses dérivés.2. Capillary affinity electrophoresis according to claim 1, characterized by the fact that the soluble polymer is polyethylene glycol (PEG) or one of its derivatives.
10 3. Support d'affinité pour électrophorèse capillaire d'affinité selon les revendications 1 ou 2, caractérisé par le fait qu'il est constitué par un ligand d'affinité couplé à un polymère soluble.3. Affinity support for capillary affinity electrophoresis according to claims 1 or 2, characterized in that it is constituted by an affinity ligand coupled to a soluble polymer.
4. Support d'affinité selon la revendication 3, caractérisé par le fait que J5 le polymère soluble est le polyéthylène glycol (PEG).4. Affinity support according to claim 3, characterized in that J5 the soluble polymer is polyethylene glycol (PEG).
5. Procédé pour la préparation du support d'affinité selon les revendi¬ cations 3 ou 4, caractérisé par le fait qu'on couple un ligand d'affinité à un poly¬ mère qui est soluble à température ambiante dans l'eau, le dichiorure de méthane,5. Process for the preparation of the affinity support according to claims 3 or 4, characterized in that an affinity ligand is coupled to a poly¬ mother which is soluble at room temperature in water, the methane dichioride,
20 l'acétonitrile et, à chaud, dans l'acétone, le dioxane, le toluène et dont la masse est supérieure à 4,996.10"24 kg et la viscosité inférieure à 5 mPa.s. 20 acetonitrile and, when hot, in acetone, dioxane, toluene and whose mass is greater than 4.996.10 "24 kg and the viscosity less than 5 mPa.s.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| FR9410928A FR2724459B1 (en) | 1994-09-14 | 1994-09-14 | AFFINITY CAPILLARY ELECTROPHORESIS, AFFINITY MATRIX FOR THE IMPLEMENTATION OF THIS ELECTROPHORESIS AND ITS MANUFACTURING METHOD |
| FR94/10928 | 1994-09-14 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1996008716A1 true WO1996008716A1 (en) | 1996-03-21 |
Family
ID=9466905
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/FR1995/001176 WO1996008716A1 (en) | 1994-09-14 | 1995-09-14 | Capillary affinity electrophoresis, affinity matrix for implementing said electrophoresis and method of manufacture |
Country Status (2)
| Country | Link |
|---|---|
| FR (1) | FR2724459B1 (en) |
| WO (1) | WO1996008716A1 (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997045721A1 (en) * | 1996-05-24 | 1997-12-04 | Novartis Ag | Process for separating mixtures of substances using capillary affinity gel electrophoresis |
| US7157546B2 (en) | 2002-09-09 | 2007-01-02 | Nektar Therapeutics Al, Corporation | Water-soluble polymer alkanals |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1500464A (en) * | 1976-03-29 | 1978-02-08 | Marine Colloids Inc | Applying reagent to molecular separation media and device therefor |
| US5108568A (en) * | 1989-07-07 | 1992-04-28 | The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration | Controlled method of reducing electrophoretic mobility of macromolecules, particles or cells |
-
1994
- 1994-09-14 FR FR9410928A patent/FR2724459B1/en not_active Expired - Fee Related
-
1995
- 1995-09-14 WO PCT/FR1995/001176 patent/WO1996008716A1/en active Application Filing
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1500464A (en) * | 1976-03-29 | 1978-02-08 | Marine Colloids Inc | Applying reagent to molecular separation media and device therefor |
| US5108568A (en) * | 1989-07-07 | 1992-04-28 | The United States Of America As Represented By The Administrator Of The National Aeronautics And Space Administration | Controlled method of reducing electrophoretic mobility of macromolecules, particles or cells |
Non-Patent Citations (1)
| Title |
|---|
| H. GOUBRAN-BOTROS: "IMMOBILIZED METAL ION AFFINITY ELECTROPHORESIS", JOURNAL OF CHROMATOGRAPHY, vol. 597, AMSTERDAM, NL, pages 357 - 364 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997045721A1 (en) * | 1996-05-24 | 1997-12-04 | Novartis Ag | Process for separating mixtures of substances using capillary affinity gel electrophoresis |
| US7157546B2 (en) | 2002-09-09 | 2007-01-02 | Nektar Therapeutics Al, Corporation | Water-soluble polymer alkanals |
| US7511094B2 (en) | 2002-09-09 | 2009-03-31 | Nektar Therapeutics Al, Corporation | Water-soluble polymer alkanals |
| US7838595B2 (en) | 2002-09-09 | 2010-11-23 | Nektar Therapeutics | Water-soluble polymer alkanals |
| US8076412B2 (en) | 2002-09-09 | 2011-12-13 | Nektar Therapeutics | Water-soluble polymer alkanals |
| US8853325B2 (en) | 2002-09-09 | 2014-10-07 | Nektar Therapeutics | Water-soluble polymer alkanals |
Also Published As
| Publication number | Publication date |
|---|---|
| FR2724459B1 (en) | 1997-05-09 |
| FR2724459A1 (en) | 1996-03-15 |
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