New strategy for the design of ligands for the purification of pharmaceutical proteins by affinity chromatography

Sproule, Kenny; Morrill, Paul; Pearson, Jennifer; Burton, Steven J.; Hejnæs, Kim R.; Valore, Henrik; Ludvigsen, Svend; Lowe, Christopher R.

doi:10.1016/s0378-4347(99)00570-8

Cited by 81 publications

(38 citation statements)

References 13 publications

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“…One of the modern suggested ways is the following. Selection of appropriate site on the target protein, design of a complementary ligand compatible with the 3D structure of the site, construction of a limited solid-phase combinatorial library of near-neighbor ligands and solution synthesis of the hit ligand, immobilization, optimization, and application of the adsorbent for the purification of the target protein [13]. This strategy is directed to the understanding of affinity and selectivity for enzyme inhibitors uses information from ligands and 3D structures of target proteins [14].…”

Section: Chemical Proteomics: Management Of Biosystems By Selective Lmentioning

confidence: 99%

Postgenomic chemistry: New problems and challenges

Varfolomeyev

Алиев

Ефременко

2004

Pure and Applied Chemistry

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New trends in chemistry induced by genetic engineering and genomic information are analyzed. Genomic information and bioinformatics make the identification of all molecular participants of biological processes possible. The methods of identification of enzymeactive sites from sequence data were demonstrated. The problems of "chemical proteomics" were formulated and analyzed. New biocatalytic processes based on a new generation of enzymes, including CO 2 reduction and enzyme fuel cell construction, are demonstrated. It was shown that in many reactions the enzymes are substantially more efficient in comparison with classical chemical catalysts. The creation of recombinant proteins from unnatural amino acids is demonstrated, and organisms containing such proteins are described. The new trends of proteomics and bioanalytical chemistry in the postgenomic era are discussed.

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Section: Chemical Proteomics: Management Of Biosystems By Selective Lmentioning

confidence: 99%

Postgenomic chemistry: New problems and challenges

Varfolomeyev

Алиев

Ефременко

2004

Pure and Applied Chemistry

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“…The ligand library was synthesized following a previously described protocol (Sproule et al, 2000), using the amines listed in Table 1. Comparison of the epoxide content (20 mmol/g moist-weight gel) with the extent of amination (18 mmol/g moist weight gel) suggested almost complete reaction of the epoxy-activated gel with ammonia.…”

Section: Rational Design and Synthesis Of The Ligand Librarymentioning

confidence: 99%

“…Used in conjunction with molecular modelling, this approach has yielded ligands for a number of different proteins, including immunoglobulin G (Li et al, 1998;Teng et al, 1999), glycoproteins (Palanisamy et al, 1999), endotoxins (Lawden et al, 1995) and trypsin-like proteases Lowe, 1992, 1993). Such ligands were generated through the synthesis of a ligand library on a solidsupport by a modified split and mix approach (Sproule et al, 2000). The ligand scaffold molecule was cyanuric chloride (2,4,6-trichloro-1,3,5-triazine), whose chlorines can be easily substituted for amines.…”

Section: Introductionmentioning

confidence: 99%

The design, synthesis and evaluation of affinity ligands for prion proteins

Renou

Gupta

Young

et al. 2004

J of Molecular Recognition

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Bifunctional affinity ligands based on a triazine scaffold were rationally designed to target prion protein and shown to bind recombinant prion protein with high affinity and selectivity. The ligands were capable of discriminating between prion protein glycoforms and monomeric and dimeric forms of the prion protein. The ligands also discriminate between conformational differences in the prion protein, resulting from point mutations in the prion protein gene. These results suggest that derived compounds could be used selectively to detect the disease-associated form of the prion protein, and as such, could provide diagnostic or therapeutic tools for prion diseases.

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“…Combinatorial methods have also facilitated the discovery of biospecific ligands for the purification of a wide variety of proteins by affinity chromatography (Burton and Lowe, 1992;Lowe, 2001;Sproule et al, 2000;Teng et al, 1999). Microtiter-plate techniques, based on immobilized metalaffinity chromatography, are routinely used for the high-throughput purification of histidine tagged proteins (Draveling et al, 2001;Paborsky et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

High-throughput process development for recombinant protein purification

Rege

Pepsin²,

Falcon³

et al. 2006

Biotechnol. Bioeng.

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Methods development in chromatographic purification processes is a complex operation and has traditionally relied on trial and error approaches. The availability of a large number of commercial media, choice of different modes of chromatography, and diverse operating conditions contribute to the challenging task of accelerating methods development. In this paper, we describe a novel microtiter-plate based screening method to identify the appropriate sequence of chromatographic steps that result in high purities of bioproducts from their respective culture broths. Protein mixtures containing the bioproduct were loaded on aliquots of different chromatographic media in microtiter plates. Serial step elution of the proteins, in concert with bioproduct-specific assays, resulted in the identification of "active fractions" containing the bioproduct. The identification of a successful chromatographic step was based on the purity of the active fractions, which were then pooled and used as starting material for screening the next chromatographic dimension. This procedure was repeated across subsequent dimensions until single band purities of the protein were obtained. The sequence of chromatographic steps and the corresponding operating conditions identified from the screen were validated under scaled-up conditions. Various modes of chromatography including hydrophobic interaction, ion exchange (cation and anion exchange) and hydrophobic charge-induction chromatography (HCIC), and different operating conditions (pH, salt concentration and type, etc.) were employed in the screen. This approach was employed to determine the sequence of chromatographic steps for the purification of recombinant alpha-amylase from its cell-free culture broth. Recommendations from the screen resulted in single-band purity of the protein under scaled-up conditions. Similar results were observed for an scFv-beta-lactamase fusion protein. The use of a miniaturized screen enables the parallel screening of a wide variety of actual bioprocess media and conditions and represents a novel paradigm approach for the high-throughput process development of recombinant proteins.

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New strategy for the design of ligands for the purification of pharmaceutical proteins by affinity chromatography

Cited by 81 publications

References 13 publications

Postgenomic chemistry: New problems and challenges

Postgenomic chemistry: New problems and challenges

The design, synthesis and evaluation of affinity ligands for prion proteins

High-throughput process development for recombinant protein purification

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