Covalent Immobilization of Proteins onto (Maleic Anhydride-<i>alt</i>-methyl Vinyl Ether) Copolymers:  Enhanced Immobilization of Recombinant Proteins

Ladavière, Catherine; Delair, Thierry; Domard, Alain; Novelli-Rousseau, A.; Mandrand, Bernard; Mallet, François

doi:10.1021/bc970208i

Cited by 36 publications

(48 citation statements)

References 14 publications

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“…For the optimized grafting conditions, the immobilization of ODN was achieved in 4 min, 6 and for proteins the required time for completion of the binding process varied from 10 to 40 min. 7,8 These data prove that the reaction of amine groups, under proper experimental conditions, can be favored over hydrolysis even in an aqueous medium. Therefore, the ratio of anhydride groups involved in the grafting reaction can be quite high, as shown, for instance, in the case of the immobilization of oligonucleotides.…”

mentioning

confidence: 58%

“…The conditions and results of coupling were reported elsewhere. 7,8 ODN can sustain a fair amount of organic solvent and, actually, coupling mixtures consisted in 5% aqueous buffer in DMSO. For proteins, because their structure is modified by organic solvents, their immobilization was achieved in aqueous mixtures containing only 5% of DMSO.…”

Section: Effect Of Physico-chemical Factors On the Immobilization Reamentioning

confidence: 99%

“…7,8 Biological molecules are only soluble in water and, even if oligonucleotides can withstand high content of organic solvent, proteins lose their biological specificities in medium containing too much organic solvent. So, for our purposes, water was a prerequisite.…”

Section: Introductionmentioning

confidence: 99%

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Covalent immobilization of biological molecules to maleic anhydride and methyl vinyl ether copolymers?A physico-chemical approach

Ladavière

Delair

Domard

et al. 1999

J. Appl. Polym. Sci.

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ABSTRACT:The covalent grafting of biological molecules to copolymers of maleic anhydride and methyl vinyl ether (MAMVE) has been used in various applications in diagnostics. To tentatively elucidate the phenomena involved in the control of the immobilization of oligodeoxynucleotides and proteins, the physico-chemical properties of MAMVE copolymers were investigated. Because the grafting mixture contains water, to allow dissolution of the biomolecules without loss of biological properties, the anhydride-based copolymer evolves from a neutral to a negatively charged macromolecule due to hydrolysis of the anhydride moities. The properties of both hydrolyzed and nonhydrolyzed polymers were investigated. As demonstrated by light-scattering measurements in batch, the copolymers showed some level of aggregation in DMF, DMSO, and aqueous DMSO. The presence of aggregates was confirmed by size-exclusion chromatography in DMF. However, partial deaggregation occurred for the lowest molecular weight sample, on adding 1% w/v of LiBr. The nonhydrolyzed copolymers exhibited a rigid conformation in a 5% water/DMSO mixture, as well as their hydrolyzed counterpart at a low ionization degree. The rate of the hydrolysis reaction was shown to be dependent on the pH of the reaction medium and on temperature. The activation energy of the hydrolysis reaction was 14 kJ/mol, and the rate constant in the order of 10 Ϫ4 s Ϫ1 . On the basis of these data, the effect on the grafting reaction of biomolecules of different parameters such as ionic strength and the nature of the solvent, along with some other results were interpreted in terms of interactions between the synthetic and bioactive macromolecules.

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mentioning

confidence: 58%

Section: Effect Of Physico-chemical Factors On the Immobilization Reamentioning

confidence: 99%

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Covalent immobilization of biological molecules to maleic anhydride and methyl vinyl ether copolymers?A physico-chemical approach

Ladavière

Delair

Domard

et al. 1999

J. Appl. Polym. Sci.

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“…[5][6][7] Many studies have employed the hydrolyzed form of the regularly alternating copolymers of maleic anhydride and alkyl vinyl ethers because of their aggregation behavior in solution. 1,4,[8][9][10][11][12][13][14] They form intramolecular nanodomains at low pH, but the extent of microdomain formation diminishes with increasing pH as a result of the repulsion between the ionized carboxylate groups. 15,16 Fluorescence quenching studies have revealed that these HM polymers form large numbers of relatively small nanodomains.…”

Section: Introductionmentioning

confidence: 99%

Conformational Changes of Pyrene-Labeled Polyelectrolytes with pH: Effect of Hydrophobic Modifications

Deo

Somasundaran

et al. 2005

J. Phys. Chem. B

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Structural changes of pyrene-labeled and unlabeled poly(maleic acid/octyl vinyl ether) (PMAOVE) and poly-(maleic acid/methyl vinyl ether) (PMAMVE) with changes in pH have been investigated in this study. The changes in the photophysical properties of pyrene are interpreted to investigate uncoiling or swelling of the polymeric chains with pH. The vibrational fine structure of the pyrene fluorescence (I 3 /I 1 ) and the ratio between excimer and monomer fluorescence (I e /I m ) of both pyrene-labeled and unlabeled PMAMVE and PMAOVE suggest that, at pH 4, the polymers are in the coiled form and PMAOVE forms hydrophobic nanodomains. An increase in pH ionizes a number of COOH groups on both PMAMVE and PMAOVE, which leads to the stretching or swelling of the polymers.

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“…As ⑀-amino groups of lysine residues of proteins spontaneously react in neutral and mildly alkaline media with the anhydride groups to form amides, maleic anhydride copolymers have already been applied for covalent protein immobilization for decades. [7][8][9] Similarly, maleic anhydride copolymers can be modified systematically by reactions of the anhydride moieties with a wide variety of functional amines. Through the choice of adequate conditions, this reaction can be restricted to a part of the anhydride groups that permits subsequent covalent protein immobilization via the remaining moieties.…”

Section: Introductionmentioning

confidence: 99%

Modification of poly(octadecene‐alt‐maleic anhydride) films by reaction with functional amines

Schmidt

Zschoche

Werner

2002

J of Applied Polymer Sci

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Thin films of poly(octadecene-alt-maleic anhydride) on top of Si wafers and glass plates were modified by reactions with different functional amines to be used in future studies on the relevance of certain molecular surface properties for the covalent immobilization of proteins. For that aim, a strategy was developed and applied to convert the anhydride moieties of the copolymer by functional amines into side chains bearing hydrophilic groups of acidic (carboxylic acid, sulfonic acid), basic (amines), or neutral (poly(ethylene oxide) (PEO), glucose) character. The modification of the copolymer films was achieved through the two-step formation of a cyclic imide, which was very stable in aqueous solution. Depending on the reactivity of the applied amine, the adjustment of the reaction time was suitable for the preparation of partially converted surfaces of the polymer film. Degrees of modification between 5 and 30% (according to X-ray photoelectron spectroscopy data) were obtained. Annealing the modified polymer films induced efficient back-formation of the anhydride groups. By reaction of the layered polyanhydrides with highly crosslinked diamines, amine-functionalized polymer films were produced that were capable of binding secondary polyanhydride layers.

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Covalent Immobilization of Proteins onto (Maleic Anhydride-alt-methyl Vinyl Ether) Copolymers: Enhanced Immobilization of Recombinant Proteins

Cited by 36 publications

References 14 publications

Covalent immobilization of biological molecules to maleic anhydride and methyl vinyl ether copolymers?A physico-chemical approach

Covalent immobilization of biological molecules to maleic anhydride and methyl vinyl ether copolymers?A physico-chemical approach

Conformational Changes of Pyrene-Labeled Polyelectrolytes with pH: Effect of Hydrophobic Modifications

Modification of poly(octadecene‐alt‐maleic anhydride) films by reaction with functional amines

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