Separation of human IgG fragments using copper, nickel, zinc, and cobalt chelated to CM-Asp-agarose by positive and negative chromatography

Mourão, Cecília Alves; Carmignotto, Gabriela Pannunzio; Bueno, Sônia Maria Alves

doi:10.1016/j.jchromb.2016.01.058

Cited by 13 publications

(21 citation statements)

References 40 publications

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“…The concentration of Fab fragments in the chromatographic fractions was determined by radial immunodiffusion (RID) assay, in accordance with da Silva et al (), Mourão et al () and Carmignotto et al (). A Fab‐specific anti‐human IgG antibody conjugated to peroxidase produced in goat was used (Sigma‐Aldrich, USA).…”

Section: Methodsmentioning

confidence: 63%

“…Because of the potential shown by P‐Tyr‐agarose for purification of IgG from human plasma, experiments on the adsorption of fragments from a papain‐digested human IgG solution were carried out. Based on the negative charges of immobilized P‐Tyr, it was possible that Fc fragments would not be adsorbed at a neutral pH (7.0), since these fragments are negatively charged at pH values above their pI range of 5.5–6.7 (Mourão et al, ).…”

Section: Resultsmentioning

confidence: 99%

“…The ligand P‐Tyr is negatively charged at pH 7.0 and the isoelectric points of the fragments are in the ranges of 6.0–9.3 and 5.5–6.7 for Fab and Fc, respectively (Mourão et al, ). At pH 7.0, Fc fragments are negatively charged, while Fab fragments have positive, negative and neutral charges.…”

Section: Resultsmentioning

confidence: 99%

“…Aiming at obtaining Fab and Fc fragments, whole human IgG was cleaved with papain following the procedures described by Ternynck and Avrameas (), da Silva, Serracchiani, Miranda, and Bueno (), Mourão, Carmignotto, and Bueno (), and Carmignotto, Mourão, and Bueno () using a 30 mg mL −1 solution of high purity human IgG in 100 mmol L −1 NaP buffer, pH 7.4 containing 40 mmol L −1 EDTA. Then cysteine‐activated papain was added to the IgG solution to achieve a papain/antibody ratio of 1% (w/w).…”

Section: Methodsmentioning

confidence: 99%

“…After SDS–PAGE runs, protein samples were transferred to nitrocellulose membranes in a Mini Transblotting III System (Bio‐Rad, USA) as described by da Silva et al (), Mourão et al () and Carmignotto et al () for the specific detection of whole human IgG and its Fc and Fab fragments. The secondary antibodies used were peroxidase conjugated anti‐human IgG (Fab specific or Fc specific), both produced in goat and purchased from Sigma‐Aldrich (USA).…”

Section: Methodsmentioning

confidence: 99%

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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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Section: Methodsmentioning

confidence: 63%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

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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Self‐assembly preparation of Zn²⁺‐immobilized silica hybrid monolith and application in solid‐phase micro‐extraction of β‐agonists

Peng

Zhang

et al. 2022

J of Separation Science

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Exploiting adsorbents with highly efficient extraction performance is of great promise for extracting small organic molecules from biological samples. In this work, a novel Zn 2+ -immobilized chitosan@silica hybrid monolith was prepared through a simple self-assembly Zn 2+ -immobilization process. Exploited as an adsorbent in solid-phase micro-extraction for extracting trace β-agonists, the monolith exhibited high extraction efficiencies for salbutamol, clenbuterol, and ractopamine with the enrichment factors approaching 120, 85, and 52, respectively. These could be attributed to the effective interaction between Zn 2+ ions and the target molecule via coordination or other intermolecular interactions. Under optimized extraction operations, a sensitive determination was successfully developed coupling with high-performance liquid chromatographyultraviolet detection. The linear range was 0.17-58.8, 0.12-68.5, and 0.18-65.5 ng/ml for salbutamol, clenbuterol, and ractopamine. The limits of detection of the β-agonists were from 0.04 to 0.07 ng/ml, and the limits of quantification were from 0.12 to 0.18 ng/ml. The recoveries of spiking in mutton samples were observed in the range of 85.9%-95.7%, with relative standard deviations <8.0%

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Rapid and regenerable surface plasmon resonance determinations of biomarker concentration and biomolecular interaction based on tris-nitrilotriacetic acid chips

Liu

Han

Jiang

et al. 2021

Analytica Chimica Acta

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Separation of human IgG fragments using copper, nickel, zinc, and cobalt chelated to CM-Asp-agarose by positive and negative chromatography

Cited by 13 publications

References 40 publications

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

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

Self‐assembly preparation of Zn²⁺‐immobilized silica hybrid monolith and application in solid‐phase micro‐extraction of β‐agonists

Rapid and regenerable surface plasmon resonance determinations of biomarker concentration and biomolecular interaction based on tris-nitrilotriacetic acid chips

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Separation of human IgG fragments using copper, nickel, zinc, and cobalt chelated to CM-Asp-agarose by positive and negative chromatography

Cited by 13 publications

References 40 publications

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

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

Self‐assembly preparation of Zn2+‐immobilized silica hybrid monolith and application in solid‐phase micro‐extraction of β‐agonists

Rapid and regenerable surface plasmon resonance determinations of biomarker concentration and biomolecular interaction based on tris-nitrilotriacetic acid chips

Contact Info

Product

Resources

About

Self‐assembly preparation of Zn²⁺‐immobilized silica hybrid monolith and application in solid‐phase micro‐extraction of β‐agonists