Design, synthesis and evaluation of pyridine-based chromatographic adsorbents for antibody purification

Mountford, Simon J.; Daly, Rachel; Robinson, Andrea J.; Hearn, Milton Thomas William

doi:10.1016/j.chroma.2014.06.006

Cited by 20 publications

(23 citation statements)

References 33 publications

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“…The 5-bromo-2-pyridinylsulfanylethylamine (5-Br-2-PSEA) was made in a similar manner from 2,5-dibromopyridine but with sodium ethoxide as the base in ethanol and microwave heating at 140°C for 21 min [22]. The source and purity of the chemicals used in these syntheses have been described in detail previously [19,20,22].…”

Section: Synthesis and Immobilization Of Pyridine-based Ligandsmentioning

confidence: 99%

“…Immobilisation of these ligands onto epichlorohydrin-activated Sepharose 6 FF TM was based on methods described previously [19,20]. The Sepharose 6 FF™ typically in aliquots of 500 mL was activated with epichlorohydrin according to described procedures [19,20].…”

Section: Synthesis and Immobilization Of Pyridine-based Ligandsmentioning

confidence: 99%

“…1) were prepared from the reaction of cysteamine hydrochloride with the appropriate aromatic chloride with sodium hydride as base [19,20]. The 5-bromo-2-pyridinylsulfanylethylamine (5-Br-2-PSEA) was made in a similar manner from 2,5-dibromopyridine but with sodium ethoxide as the base in ethanol and microwave heating at 140°C for 21 min [22].…”

Section: Synthesis and Immobilization Of Pyridine-based Ligandsmentioning

confidence: 99%

“…The structures of our pyridine-based ligands, when immobilised onto various support materials, allow similar mixed mode binding behaviour to be achieved, due to the participation of electrostatic and thiophilic-based interactions depending on the compositions and pH of the chosen loading, wash and elution buffers. Interestingly, in preliminary studies chromatographic resins generated from the immobilisation of several of these compounds showed high binding capacities for mAbs, which could be successfully eluted in a highly efficient manner through a step change in buffer pH [19,20].…”

Section: Introductionmentioning

confidence: 99%

“…Generally, mixed mode ligands can be grouped into three subsets, based on the characteristics of the dominant functionalities: (1) those augmenting anion exchange behaviour with hydrogen bonding effects; (2) those augmenting metal coordination with electrostatic interactions; and (3) those augmenting hydrophobic interactions with dipole-dipole or van der Waals characteristics [6,12]. http In previous communications, we have described the synthesis of different libraries of pyridine-based compounds and evaluated some examples for their potential as ligands in the affinity and mixed mode chromatographic purification of recombinant proteins [19,20]. These ligands, comprising a pyridine core with a variety of different substituent moieties, have also included analogues with one or more additional aromatic ring structures which show increased protein binding attributes.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Zhang

Fredericks

Campi

et al. 2015

Separation and Purification Technology

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Section: Synthesis and Immobilization Of Pyridine-based Ligandsmentioning

confidence: 99%

Section: Synthesis and Immobilization Of Pyridine-based Ligandsmentioning

confidence: 99%

Section: Synthesis and Immobilization Of Pyridine-based Ligandsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Zhang

Fredericks

Campi

et al. 2015

Separation and Purification Technology

Self Cite

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QSAR models for prediction of chromatographic behavior of homologous Fab variants

Robinson

Karkov

Woo

et al. 2017

Biotech & Bioengineering

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While quantitative structure activity relationship (QSAR) models have been employed successfully for the prediction of small model protein chromatographic behavior, there have been few reports to date on the use of this methodology for larger, more complex proteins. Recently our group generated focused libraries of antibody Fab fragment variants with different combinations of surface hydrophobicities and electrostatic potentials, and demonstrated that the unique selectivities of multimodal resins can be exploited to separate these Fab variants. In this work, results from linear salt gradient experiments with these Fabs were employed to develop QSAR models for six chromatographic systems, including multimodal (Capto MMC, Nuvia cPrime, and two novel ligand prototypes), hydrophobic interaction chromatography (HIC; Capto Phenyl), and cation exchange (CEX; CM Sepharose FF) resins. The models utilized newly developed "local descriptors" to quantify changes around point mutations in the Fab libraries as well as novel cluster descriptors recently introduced by our group. Subsequent rounds of feature selection and linearized machine learning algorithms were used to generate robust, well-validated models with high training set correlations (R > 0.70) that were well suited for predicting elution salt concentrations in the various systems. The developed models then were used to predict the retention of a deamidated Fab and isotype variants, with varying success. The results represent the first successful utilization of QSAR for the prediction of chromatographic behavior of complex proteins such as Fab fragments in multimodal chromatographic systems. The framework presented here can be employed to facilitate process development for the purification of biological products from product-related impurities by in silico screening of resin alternatives. Biotechnol. Bioeng. 2017;114: 1231-1240. © 2016 Wiley Periodicals, Inc.

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Homo‐ and Heterodinuclear Rh and Ir Complexes Supported by SN_n Mixed‐Donor Ligands (n = 2–4): Stereochemistry and Coordination‐Site‐Exchange Reactions of Cp*M (M = Rh, Ir) Units

et al. 2016

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A series of SNn mixed‐donor ligands [n = 2: H2NC2H4SCH2‐2‐pyridyl (2‐NSpy) (1a), H2NC2H4SCH2‐4‐pyridyl (4‐NSpy) (1b), n = 3: 2‐pyridylCH2NHC2H4SCH2‐2‐pyridyl (2‐pyNSpy) (2), n = 4: (2‐pyridylCH2)2NC2H4SCH2‐2‐pyridyl (2‐py2NSpy) (3)] was utilized to support homo‐ and heterodinuclear complexes including Cp*MIII units (M = Rh, Ir; Cp* = pentamethylcyclopentadienyl). Reactions of [Cp*MCl2]2 with 2‐pyNSpy (2), 2‐py2NSpy (3), and 4‐NSpy (1b) afforded homodinulear complexes, [(Cp*MCl)(2‐pyNSpy)(Cp*MCl)](PF6)2 [M = Rh (5a), Ir (5b)], [(Cp*M)(2‐py2NSpy)(Cp*MCl)](PF6)3 [M = Rh (6a), Ir (6b)], [(Cp*MCl)(4‐NSpy)(Cp*MCl2)]Cl [M = Rh (8a), Ir (8b)]. Heterodinuclear complexes [(Cp*MCl)(4‐NSpy)(Cp*M′Cl2)]Cl [M, M′ = Rh, Ir (8c), Ir, Rh (8d)] were prepared using mononuclear complexes [(Cp*MCl)(4‐NSpy)]Cl [M = Rh (7a), Ir (7b)] reacted with [Cp*MCl2]2 (M = Ir, Rh), respectively. Complexes 5–8 were characterized by X‐ray crystallography to determine the configurations around the M, M′, S, and N centers. The solid‐state structures of 6 are retained in acetonitrile solution whereas four diastereomers are generated in the case of 5 due to low stereoselectivity around the coordinated amine nitrogen atom, in contrast to the sulfur atom. Heterodinuclear complexes 8c,d are unstable in solution at 55 °C, readily affording mixtures of 8a–d via intra‐ and intermolecular coordination‐site‐exchange reactions of Cp*M fragments between the SN moiety and the py site. In order to evaluate the selectivity of Cp*M fragments for the SN and py coordination sites, several competitive reactions of [Cp*MCl2]2 (M = Rh, Ir) with H2NC2H4SCH2C6H5 (NSph) (4) and/or 4‐methylpyridine (4‐Mepy) were carried out to demonstrate predominant formation of iridium complexes 9b and 10b among [(Cp*MCl)(NSph)]Cl [M = Rh (9a), Ir (9b)] and [(Cp*MCl)(4‐Mepy)]Cl [M = Rh (10a), Ir (10b)]. These reactions indicated higher affinity of the Cp*Ir fragment to both the NS and py sites relative to the rhodium analogue.

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Design, synthesis and evaluation of pyridine-based chromatographic adsorbents for antibody purification

Cited by 20 publications

References 33 publications

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

QSAR models for prediction of chromatographic behavior of homologous Fab variants

Homo‐ and Heterodinuclear Rh and Ir Complexes Supported by SN_n Mixed‐Donor Ligands (n = 2–4): Stereochemistry and Coordination‐Site‐Exchange Reactions of Cp*M (M = Rh, Ir) Units

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Design, synthesis and evaluation of pyridine-based chromatographic adsorbents for antibody purification

Cited by 20 publications

References 33 publications

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

Purification of monoclonal antibodies by chemical affinity mixed mode chromatography

QSAR models for prediction of chromatographic behavior of homologous Fab variants

Homo‐ and Heterodinuclear Rh and Ir Complexes Supported by SNn Mixed‐Donor Ligands (n = 2–4): Stereochemistry and Coordination‐Site‐Exchange Reactions of Cp*M (M = Rh, Ir) Units

Contact Info

Product

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Homo‐ and Heterodinuclear Rh and Ir Complexes Supported by SN_n Mixed‐Donor Ligands (n = 2–4): Stereochemistry and Coordination‐Site‐Exchange Reactions of Cp*M (M = Rh, Ir) Units