Molecular insight in the purification of immunoglobulin by pseudobiospecific ligand l-histidine and histidyl moieties in histidine ligand affinity chromatography (HLAC) by molecular docking

Savane, Tushar S.; Kumar, Sanjit; Janakiraman, Vignesh Narasimhan; Kamalanathan, A.S.; Vijayalakshmi, M.A.

doi:10.1016/j.jchromb.2015.09.010

Cited by 11 publications

(5 citation statements)

References 39 publications

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“…Numerous studies have investigated the effect of the physicochemical properties of the spacer arm (length, flexibility, and hydrophilicity) on the binding capacity and selectivity of the adsorbent [51,52,53]. The optimization of the spacer arm has been consistently found to improve the ligand display on the surface of the resin, and therefore the performance of the affinity adsorbent [35,36,54,55,56,57].…”

Section: Resultsmentioning

confidence: 99%

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Islam

Naik

Hashimoto

et al. 2019

IJMS

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This work presents the use of peptide ligand HWRGWV and its cognate sequences to develop affinity adsorbents that compete with Protein A in terms of binding capacity and quality of the eluted product. First, the peptide ligand was conjugated to crosslinked agarose resins (WorkBeads) at different densities and using different spacer arms. The optimization of ligand density and display resulted in values of static and dynamic binding capacity of 85 mg/mL and 65 mg/mL, respectively. A selected peptide-WorkBeads adsorbent was utilized for purifying Mabs from Chinese Hamster Ovary (CHO) cell culture supernatants. The peptide-WorkBeads adsorbent was found able to withstand sanitization with strong alkaline solutions (0.5 M NaOH). The purity of the eluted product was consistently higher than 95%, with logarithmic removal value (LRV) of 1.5 for host cell proteins (HCPs) and 4.0 for DNA. HCP clearance was significantly improved by adding a post-load washing step with either 0.1 M Tris HCl pH 9 or 1 M NaCl. The cognate peptide of HWRGWV, constructed by replacing arginine (R) with citrulline, further increased the HCP LRV to 2.15. The peptide-based adsorbent also showed a remarkable performance in terms of removal of Mab aggregates; unlike Protein A, in fact, HWRGWV was found to bind only monomeric IgG. Collectively, these results demonstrate the potential of peptide-based adsorbents as alternative to Protein A for the purification of therapeutic antibodies.

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Section: Resultsmentioning

confidence: 99%

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Islam

Naik

Hashimoto

et al. 2019

IJMS

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“…However, the binding specificity of IgG molecules on immobilized histidine depends on the nature of the matrix, ligand orientation and chromatographic conditions. The interactions between histidine and IgG are predominantly electrostatic, but others such as mild hydrophobic, van der Waals and hydrogen bonding play an important role (Prasanna et al, ; Savane, Kumar, Janakiraman, Kamalanathan, & Vijayalakshmi, ). Immunoglobulins are adsorbed onto immobilized histidine at low ionic strength and eluted under mild conditions and the purity of the isolated IgG reaches ~90%.…”

Section: Introductionmentioning

confidence: 99%

Phospho‐l‐tyrosine‐agarose chromatography: Adsorption of human IgG and its proteolytic fragments

Pavan

Bresolin

Muzio

et al. 2018

Biomedical Chromatography

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The behavior of human immunoglobulin G (IgG) and antigen-binding fragment (Fab fragment) adsorption onto phospho-L-tyrosine immobilized on agarose (P-Tyr-agarose) was evaluated by pseudoaffinity chromatography. The effects of buffer systems MES, MOPS, Bis-Tris, Tris-HCl and sodium phosphate (NaP) and pH on IgG adsorption were studied and high purity values were obtained (96%, based on ELISA analysis of albumin, transferrin and immunoglobulins A, G and M) when IgG was purified from human plasma diluted in 10 mmol L −1 NaP buffer at pH 6.0. The capture of IgG by the P-Tyragarose was also promising, since 91% of the IgG was adsorbed when plasma was diluted in 25 mmol L −1 MES buffer at pH 5.5, recommending its use for IgG depletion from human plasma under this condition. The experimental data on IgG adsorption kinetics were in agreement with the pseudo-second-order model. The adsorption isotherm data were well described by the Langmuir-Freundlich model with the value of parameter n being <1 (0.72), indicating negative cooperativity. Selectivity was achieved on P-Tyr-agarose from digested human IgG in HEPES 25 mmol L −1 buffer at pH 7.0where Fab fragments were obtained in eluted fractions without Fc fragments (but with uncleaved IgG) with 86.2% recovery.

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“…Os ligantes chamados de pseudobioespecíficos, por sua vez, podem ou não ser moléculas biológicas. Como exemplos de moléculas biológicas dessa classe de ligantes, destacam-se os peptídeos e aminoácidos; exemplos de moléculas não biológicas são corantes, quelatos metálicos, aminas, tióis e boronatos (SAVANE et al, 2016).…”

Section: Tipos De Ligantes Empregados Em Cromatografia De Afinidadeunclassified

“…Nesse contexto, ao otimizar algumas propriedades da cromatografia de afinidade como o suporte sólido, o tipo de ligante e as condições a serem empregadas, a técnica pode ser aplicada à purificação em larga escala, atingindo altos rendimentos, com nível elevado de pureza (ARORA, SAXENA e AYYAR, 2017).Atualmente, a literatura traz alguns ligantes pseudobioespecíficos capazes de capturar IgG de forma seletiva. No geral, esses ligantes compreendem moléculas menores, robustas e de fácil imobilização, as quais apresentam menor custo de obtenção e maior estabilidade química e física durante os processos de sanitização e esterilização(VIJAYALAKSMI, 1989;SAVANE et al, 2016). Dentre os ligantes pseudobioespecíficos capazes de purificar IgG, destacam-se os aminoácidos como histidina, fenilalanina, arginina, triptofano, o-fosfo-L-serina e o-fosfo-L-tirosina, imobilizados em diferentes suporte sólidos(HWANG e PARK, 2003;ELKAK et al, 2009; NAIK, RAINA e LALI, 2011; BRESOLIN e BUENO, 2012;PAVAN et al, 2017).…”

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Síntese e avaliação antitumoral "in vitro" de híbridos da goniotalamina-monastrol e piplartina-monastrol

Marcuz¹

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Agradeço primeiramente a Deus, por ser presente em minha vida. Aos meus paisNeusa de Souza Baggio Marcuz e Marcelo Marcuz, por serem os responsáveis pela pessoa que me tornei. Por terem me criado com amor e aceitarem todas as minhas escolhas. Sou eternamente grata pela educação que me deram e por me incentivarem a jamais desistir. Amo vocês. Ao meu amado esposo e parceiro Caio dos Santos, por ser dono do abraço que me conforta e responsável pelo amor que me fortalece. Agradeço-lhe pelo amor, amizade e companheirismo. Não imagino a vida sem você ao meu lado. À professora Dra. Sônia Maria Alves Bueno, não só pela orientação e ensinamentos, mas também pelo empenho e dedicação profissional. Agradeço pelos conselhos e por se preocupar comigo, além da pesquisa. Aos professores Dra. Ângela Maria Moraes e Dr. Everson Alves Miranda, por disponibilizarem as instalações de seus laboratórios. Ao professor Dr. Everson Alves Miranda e à Dra. Gisele Luiza Pavan Pereira, pelo tempo dedicado a leitura do presente trabalho. Aos colegas e ex-colegas de laboratório Gisele Pavan, Cecília Mourão, Daniele Cunha, Gabriela Nascimento e Igor Fioravante, eu aprendi muito com vocês. Agradeço também aos demais colegas que compartilham as mesmas instalações:

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Molecular insight in the purification of immunoglobulin by pseudobiospecific ligand l-histidine and histidyl moieties in histidine ligand affinity chromatography (HLAC) by molecular docking

Cited by 11 publications

References 39 publications

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Phospho‐l‐tyrosine‐agarose chromatography: Adsorption of human IgG and its proteolytic fragments

Síntese e avaliação antitumoral "in vitro" de híbridos da goniotalamina-monastrol e piplartina-monastrol

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